PMC:1942070 / 0-4137
Annnotations
2_test
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| 17349631-10859332-30580518 | 1348-1349 | 10859332 | denotes | 3 |
| 17349631-12506120-30580518 | 1348-1349 | 12506120 | denotes | 3 |
| 17349631-12506120-30580519 | 1662-1663 | 12506120 | denotes | 6 |
| 17349631-9765302-30580519 | 1662-1663 | 9765302 | denotes | 6 |
| 17349631-11062248-30580519 | 1662-1663 | 11062248 | denotes | 6 |
| 17349631-11410586-30580519 | 1662-1663 | 11410586 | denotes | 6 |
| 17349631-15590638-30580520 | 1807-1808 | 15590638 | denotes | 7 |
| 17349631-10471840-30580521 | 1884-1886 | 10471840 | denotes | 12 |
| 17349631-10856238-30580521 | 1884-1886 | 10856238 | denotes | 12 |
| 17349631-11410587-30580521 | 1884-1886 | 11410587 | denotes | 12 |
| 17349631-12646243-30580521 | 1884-1886 | 12646243 | denotes | 12 |
| 17349631-12676944-30580521 | 1884-1886 | 12676944 | denotes | 12 |
| 17349631-11912133-30580522 | 1994-1996 | 11912133 | denotes | 16 |
| 17349631-11514571-30580522 | 1994-1996 | 11514571 | denotes | 16 |
| 17349631-16899224-30580522 | 1994-1996 | 16899224 | denotes | 16 |
| 17349631-12505989-30580523 | 2026-2028 | 12505989 | denotes | 18 |
| 17349631-14654790-30580523 | 2026-2028 | 14654790 | denotes | 18 |
| 17349631-14563314-30580524 | 2057-2059 | 14563314 | denotes | 19 |
| 17349631-16678913-30580525 | 2218-2220 | 16678913 | denotes | 20 |
| 17349631-12505989-30580526 | 2382-2384 | 12505989 | denotes | 23 |
| 17349631-15604256-30580526 | 2382-2384 | 15604256 | denotes | 23 |
| 17349631-15226414-30580526 | 2382-2384 | 15226414 | denotes | 23 |
| 17349631-15728188-30580527 | 2438-2440 | 15728188 | denotes | 24 |
| 17349631-16875491-30580528 | 2527-2529 | 16875491 | denotes | 25 |
| 17349631-16449666-30580529 | 2708-2710 | 16449666 | denotes | 28 |
| 17349631-15738054-30580529 | 2708-2710 | 15738054 | denotes | 28 |
| 17349631-15623513-30580529 | 2708-2710 | 15623513 | denotes | 28 |
| 17349631-15367659-30580529 | 2708-2710 | 15367659 | denotes | 28 |
| 17349631-16449666-30580530 | 2855-2856 | 16449666 | denotes | 1 |
| 17349631-16449666-30580531 | 3155-3156 | 16449666 | denotes | 1 |
| 17349631-16449666-30580532 | 3922-3923 | 16449666 | denotes | 1 |
| 17349631-16449666-30580533 | 4035-4036 | 16449666 | denotes | 1 |
| 17349631-7730364-30580534 | 4113-4115 | 7730364 | denotes | 29 |
MyTest
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| 17349631-10859332-30580518 | 1348-1349 | 10859332 | denotes | 3 |
| 17349631-12506120-30580518 | 1348-1349 | 12506120 | denotes | 3 |
| 17349631-12506120-30580519 | 1662-1663 | 12506120 | denotes | 6 |
| 17349631-9765302-30580519 | 1662-1663 | 9765302 | denotes | 6 |
| 17349631-11062248-30580519 | 1662-1663 | 11062248 | denotes | 6 |
| 17349631-11410586-30580519 | 1662-1663 | 11410586 | denotes | 6 |
| 17349631-15590638-30580520 | 1807-1808 | 15590638 | denotes | 7 |
| 17349631-10471840-30580521 | 1884-1886 | 10471840 | denotes | 12 |
| 17349631-10856238-30580521 | 1884-1886 | 10856238 | denotes | 12 |
| 17349631-11410587-30580521 | 1884-1886 | 11410587 | denotes | 12 |
| 17349631-12646243-30580521 | 1884-1886 | 12646243 | denotes | 12 |
| 17349631-12676944-30580521 | 1884-1886 | 12676944 | denotes | 12 |
| 17349631-11912133-30580522 | 1994-1996 | 11912133 | denotes | 16 |
| 17349631-11514571-30580522 | 1994-1996 | 11514571 | denotes | 16 |
| 17349631-16899224-30580522 | 1994-1996 | 16899224 | denotes | 16 |
| 17349631-12505989-30580523 | 2026-2028 | 12505989 | denotes | 18 |
| 17349631-14654790-30580523 | 2026-2028 | 14654790 | denotes | 18 |
| 17349631-14563314-30580524 | 2057-2059 | 14563314 | denotes | 19 |
| 17349631-16678913-30580525 | 2218-2220 | 16678913 | denotes | 20 |
| 17349631-12505989-30580526 | 2382-2384 | 12505989 | denotes | 23 |
| 17349631-15604256-30580526 | 2382-2384 | 15604256 | denotes | 23 |
| 17349631-15226414-30580526 | 2382-2384 | 15226414 | denotes | 23 |
| 17349631-15728188-30580527 | 2438-2440 | 15728188 | denotes | 24 |
| 17349631-16875491-30580528 | 2527-2529 | 16875491 | denotes | 25 |
| 17349631-16449666-30580529 | 2708-2710 | 16449666 | denotes | 28 |
| 17349631-15738054-30580529 | 2708-2710 | 15738054 | denotes | 28 |
| 17349631-15623513-30580529 | 2708-2710 | 15623513 | denotes | 28 |
| 17349631-15367659-30580529 | 2708-2710 | 15367659 | denotes | 28 |
| 17349631-16449666-30580530 | 2855-2856 | 16449666 | denotes | 1 |
| 17349631-16449666-30580531 | 3155-3156 | 16449666 | denotes | 1 |
| 17349631-16449666-30580532 | 3922-3923 | 16449666 | denotes | 1 |
| 17349631-16449666-30580533 | 4035-4036 | 16449666 | denotes | 1 |
| 17349631-7730364-30580534 | 4113-4115 | 7730364 | denotes | 29 |
pmc-enju-pas
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T49 | 180-187 | NN | denotes | protein |
| T50 | 188-194 | NN | denotes | kinase |
| T51 | 195-196 | NN | denotes | D |
| T52 | 197-198 | -LRB- | denotes | ( |
| T53 | 198-201 | NN | denotes | PKD |
| T54 | 201-202 | -RRB- | denotes | ) |
| T55 | 203-210 | NN | denotes | enzymes |
| T56 | 211-213 | PRP | denotes | we |
| T57 | 214-223 | VB | denotes | generated |
| T58 | 224-225 | DT | denotes | a |
| T59 | 226-234 | JJ | denotes | PKD-null |
| T60 | 235-239 | NN | denotes | DT40 |
| T61 | 240-252 | NN | denotes | B-lymphocyte |
| T62 | 253-257 | NN | denotes | cell |
| T63 | 258-262 | NN | denotes | line |
| T64 | 264-274 | RB | denotes | Previously |
| T65 | 275-277 | PRP | denotes | we |
| T66 | 278-282 | VB | denotes | have |
| T67 | 283-288 | VB | denotes | shown |
| T68 | 289-293 | IN | denotes | that |
| T69 | 294-298 | NN | denotes | PKDs |
| T70 | 299-303 | VB | denotes | have |
| T71 | 304-306 | DT | denotes | an |
| T72 | 307-316 | JJ | denotes | essential |
| T73 | 317-321 | NN | denotes | role |
| T74 | 322-324 | IN | denotes | in |
| T75 | 325-335 | VB | denotes | regulating |
| T76 | 336-341 | NN | denotes | class |
| T77 | 342-344 | CD | denotes | II |
| T78 | 345-352 | NN | denotes | histone |
| T79 | 353-365 | NN | denotes | deacetylases |
| T80 | 366-368 | IN | denotes | in |
| T81 | 369-373 | NN | denotes | DT40 |
| T82 | 374-381 | NN | denotes | B-cells |
| T83 | 382-383 | -LRB- | denotes | [ |
| T84 | 383-391 | NNP | denotes | Matthews |
| T85 | 391-392 | -COMMA- | denotes | , |
| T86 | 393-397 | NNP | denotes | S.A. |
| T87 | 397-398 | -COMMA- | denotes | , |
| T88 | 399-402 | NNP | denotes | Liu |
| T89 | 402-403 | -COMMA- | denotes | , |
| T90 | 404-406 | NNP | denotes | P. |
| T91 | 406-407 | -COMMA- | denotes | , |
| T92 | 408-416 | NNP | denotes | Spitaler |
| T93 | 416-417 | -COMMA- | denotes | , |
| T94 | 418-420 | NNP | denotes | M. |
| T95 | 420-421 | -COMMA- | denotes | , |
| T96 | 422-427 | NNP | denotes | Olson |
| T97 | 427-428 | -COMMA- | denotes | , |
| T98 | 429-433 | NNP | denotes | E.N. |
| T99 | 433-434 | -COMMA- | denotes | , |
| T100 | 435-443 | NNP | denotes | McKinsey |
| T101 | 443-444 | -COMMA- | denotes | , |
| T102 | 445-449 | NNP | denotes | T.A. |
| T103 | 449-450 | -COMMA- | denotes | , |
| T104 | 451-459 | NNP | denotes | Cantrell |
| T105 | 459-460 | -COMMA- | denotes | , |
| T106 | 461-465 | NNP | denotes | D.A. |
| T107 | 466-469 | CC | denotes | and |
| T108 | 470-481 | NNP | denotes | Scharenberg |
| T109 | 481-482 | -COMMA- | denotes | , |
| T110 | 483-487 | NNP | denotes | A.M. |
| T111 | 488-489 | -LRB- | denotes | ( |
| T112 | 489-493 | CD | denotes | 2006 |
| T113 | 493-494 | -RRB- | denotes | ) |
| T114 | 495-504 | JJ | denotes | Essential |
| T115 | 505-509 | NN | denotes | role |
| T116 | 510-513 | IN | denotes | for |
| T117 | 514-521 | NN | denotes | protein |
| T118 | 522-528 | NN | denotes | kinase |
| T119 | 529-530 | NN | denotes | D |
| T120 | 531-537 | NN | denotes | family |
| T121 | 538-545 | NN | denotes | kinases |
| T122 | 546-548 | IN | denotes | in |
| T123 | 549-552 | DT | denotes | the |
| T124 | 553-563 | NN | denotes | regulation |
| T125 | 564-566 | IN | denotes | of |
| T126 | 567-572 | NN | denotes | class |
| T127 | 573-575 | CD | denotes | II |
| T128 | 576-583 | NN | denotes | histone |
| T129 | 584-596 | NN | denotes | deacetylases |
| T130 | 597-599 | IN | denotes | in |
| T131 | 600-601 | NN | denotes | B |
| T132 | 602-613 | NN | denotes | lymphocytes |
| T133 | 615-619 | NNP | denotes | Mol. |
| T134 | 620-624 | NNP | denotes | Cell |
| T135 | 625-630 | NNP | denotes | Biol. |
| T136 | 631-633 | CD | denotes | 26 |
| T137 | 633-634 | -COMMA- | denotes | , |
| T138 | 635-644 | CD | denotes | 1569–1577 |
| T139 | 644-645 | -RRB- | denotes | ] |
| T140 | 647-649 | PRP | denotes | We |
| T141 | 650-653 | RB | denotes | now |
| T142 | 654-658 | VB | denotes | show |
| T143 | 659-663 | IN | denotes | that |
| T144 | 664-668 | NN | denotes | PKDs |
| T145 | 669-672 | VB | denotes | are |
| T146 | 673-677 | RB | denotes | also |
| T147 | 678-686 | VB | denotes | required |
| T148 | 687-689 | TO | denotes | to |
| T149 | 690-698 | VB | denotes | regulate |
| T150 | 699-704 | NN | denotes | HSP27 |
| T151 | 705-720 | NN | denotes | phosphorylation |
| T152 | 721-723 | IN | denotes | in |
| T153 | 724-728 | NN | denotes | DT40 |
| T154 | 729-736 | NN | denotes | B-cells |
| T155 | 738-745 | RB | denotes | However |
| T156 | 745-746 | -COMMA- | denotes | , |
| T157 | 747-749 | IN | denotes | in |
| T158 | 750-758 | NN | denotes | contrast |
| T159 | 759-761 | TO | denotes | to |
| T160 | 762-770 | JJ | denotes | previous |
| T161 | 771-783 | NN | denotes | observations |
| T162 | 784-786 | IN | denotes | in |
| T163 | 787-792 | JJ | denotes | other |
| T164 | 793-797 | NN | denotes | cell |
| T165 | 798-803 | NN | denotes | types |
| T166 | 803-804 | -COMMA- | denotes | , |
| T167 | 805-808 | NN | denotes | PKD |
| T168 | 809-816 | NN | denotes | enzymes |
| T169 | 817-819 | VB | denotes | do |
| T170 | 820-823 | RB | denotes | not |
| T171 | 824-832 | VB | denotes | regulate |
| T172 | 833-838 | JJ | denotes | basic |
| T173 | 839-847 | JJ | denotes | cellular |
| T174 | 848-857 | NN | denotes | processes |
| T175 | 858-862 | JJ | denotes | such |
| T176 | 863-865 | IN | denotes | as |
| T177 | 866-879 | NN | denotes | proliferation |
| T178 | 880-882 | CC | denotes | or |
| T179 | 883-891 | NN | denotes | survival |
| T180 | 892-901 | NN | denotes | responses |
| T181 | 901-902 | -COMMA- | denotes | , |
| T182 | 903-906 | CC | denotes | nor |
| T183 | 907-911 | NN | denotes | NFκB |
| T184 | 912-927 | JJ | denotes | transcriptional |
| T185 | 928-936 | NN | denotes | activity |
| T186 | 937-947 | RB | denotes | downstream |
| T187 | 948-950 | IN | denotes | of |
| T188 | 951-954 | DT | denotes | the |
| T189 | 955-956 | NN | denotes | B |
| T190 | 957-961 | NN | denotes | cell |
| T191 | 962-969 | NN | denotes | antigen |
| T192 | 970-978 | NN | denotes | receptor |
| T193 | 980-984 | RB | denotes | Thus |
| T194 | 984-985 | -COMMA- | denotes | , |
| T195 | 986-990 | NN | denotes | PKDs |
| T196 | 991-995 | VB | denotes | have |
| T197 | 996-997 | DT | denotes | a |
| T198 | 998-1007 | JJ | denotes | selective |
| T199 | 1008-1012 | NN | denotes | role |
| T200 | 1013-1015 | IN | denotes | in |
| T201 | 1016-1020 | NN | denotes | DT40 |
| T202 | 1021-1027 | NN | denotes | B-cell |
| T203 | 1028-1035 | NN | denotes | biology |
| T532 | 635-1057 | NN | denotes | 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The |
| T533 | 1058-1065 | NN | denotes | protein |
| T534 | 1066-1072 | NN | denotes | kinase |
| T535 | 1073-1074 | NN | denotes | D |
| T536 | 1075-1076 | -LRB- | denotes | ( |
| T537 | 1076-1079 | NN | denotes | PKD |
| T538 | 1079-1080 | -RRB- | denotes | ) |
| T539 | 1081-1097 | NN | denotes | serine/threonine |
| T540 | 1098-1104 | NN | denotes | kinase |
| T541 | 1105-1111 | NN | denotes | family |
| T542 | 1112-1115 | VB | denotes | has |
| T543 | 1116-1121 | CD | denotes | three |
| T544 | 1122-1129 | NN | denotes | members |
| T545 | 1129-1130 | -COLON- | denotes | : |
| T546 | 1131-1135 | NN | denotes | PKD1 |
| T547 | 1135-1136 | -COMMA- | denotes | , |
| T548 | 1137-1141 | NN | denotes | PKD2 |
| T549 | 1142-1145 | CC | denotes | and |
| T550 | 1146-1150 | NN | denotes | PKD3 |
| T551 | 1152-1156 | JJ | denotes | Most |
| T552 | 1157-1161 | NN | denotes | cell |
| T553 | 1162-1167 | NN | denotes | types |
| T554 | 1168-1175 | VB | denotes | express |
| T555 | 1176-1178 | IN | denotes | at |
| T556 | 1179-1184 | JJ | denotes | least |
| T557 | 1185-1188 | CD | denotes | two |
| T558 | 1189-1192 | NN | denotes | PKD |
| T559 | 1193-1201 | NN | denotes | isoforms |
| T560 | 1202-1205 | CC | denotes | but |
| T561 | 1206-1209 | NN | denotes | PKD |
| T562 | 1210-1217 | NN | denotes | enzymes |
| T563 | 1218-1221 | VB | denotes | are |
| T564 | 1222-1232 | RB | denotes | especially |
| T565 | 1233-1239 | RB | denotes | highly |
| T566 | 1240-1249 | VB | denotes | expressed |
| T567 | 1250-1252 | IN | denotes | in |
| T568 | 1253-1267 | JJ | denotes | haematopoietic |
| T569 | 1268-1273 | NN | denotes | cells |
| T570 | 1273-1274 | -COMMA- | denotes | , |
| T571 | 1275-1280 | WRB | denotes | where |
| T572 | 1281-1285 | PRP | denotes | they |
| T573 | 1286-1289 | VB | denotes | are |
| T574 | 1290-1299 | VB | denotes | activated |
| T575 | 1300-1302 | IN | denotes | in |
| T576 | 1303-1311 | NN | denotes | response |
| T577 | 1312-1314 | TO | denotes | to |
| T578 | 1315-1322 | NN | denotes | antigen |
| T579 | 1323-1332 | NN | denotes | receptors |
| T580 | 1333-1344 | NN | denotes | stimulation |
| T581 | 1345-1346 | -LRB- | denotes | [ |
| T582 | 1346-1349 | CD | denotes | 2,3 |
| T583 | 1349-1350 | -RRB- | denotes | ] |
| T584 | 1352-1353 | DT | denotes | A |
| T585 | 1354-1363 | VB | denotes | conserved |
| T586 | 1364-1374 | NN | denotes | signalling |
| T587 | 1375-1382 | NN | denotes | pathway |
| T588 | 1383-1390 | VB | denotes | linking |
| T589 | 1391-1398 | NN | denotes | antigen |
| T590 | 1399-1408 | NN | denotes | receptors |
| T591 | 1409-1411 | TO | denotes | to |
| T592 | 1412-1416 | NN | denotes | PKDs |
| T593 | 1417-1425 | VB | denotes | involves |
| T594 | 1426-1429 | DT | denotes | the |
| T595 | 1430-1440 | NN | denotes | activation |
| T596 | 1441-1443 | IN | denotes | of |
| T597 | 1444-1448 | NN | denotes | PLCγ |
| T598 | 1449-1452 | CC | denotes | and |
| T599 | 1453-1456 | DT | denotes | the |
| T600 | 1457-1467 | JJ | denotes | subsequent |
| T601 | 1468-1478 | NN | denotes | production |
| T602 | 1479-1481 | IN | denotes | of |
| T603 | 1482-1496 | NN | denotes | diacylglycerol |
| T604 | 1497-1498 | -LRB- | denotes | ( |
| T605 | 1498-1501 | NN | denotes | DAG |
| T606 | 1501-1502 | -RRB- | denotes | ) |
| T607 | 1503-1508 | WDT | denotes | which |
| T608 | 1509-1519 | VB | denotes | stimulates |
| T609 | 1520-1529 | JJ | denotes | classical |
| T610 | 1530-1536 | CC | denotes | and/or |
| T611 | 1537-1542 | JJ | denotes | novel |
| T612 | 1543-1550 | NN | denotes | protein |
| T613 | 1551-1557 | NN | denotes | kinase |
| T614 | 1558-1560 | NN | denotes | Cs |
| T615 | 1561-1562 | -LRB- | denotes | ( |
| T616 | 1562-1565 | NN | denotes | PKC |
| T617 | 1565-1566 | -RRB- | denotes | ) |
| T618 | 1567-1571 | WDT | denotes | that |
| T619 | 1572-1585 | VB | denotes | phosphorylate |
| T620 | 1586-1589 | CD | denotes | two |
| T621 | 1590-1593 | JJ | denotes | key |
| T622 | 1594-1604 | JJ | denotes | regulatory |
| T623 | 1605-1611 | NN | denotes | serine |
| T624 | 1612-1620 | NN | denotes | residues |
| T625 | 1621-1623 | IN | denotes | in |
| T626 | 1624-1627 | DT | denotes | the |
| T627 | 1628-1638 | NN | denotes | activation |
| T628 | 1639-1643 | NN | denotes | loop |
| T629 | 1644-1646 | IN | denotes | of |
| T630 | 1647-1650 | NN | denotes | PKD |
| T631 | 1651-1658 | NN | denotes | kinases |
| T632 | 1659-1660 | -LRB- | denotes | [ |
| T633 | 1660-1663 | CD | denotes | 3–6 |
| T634 | 1663-1664 | -RRB- | denotes | ] |
| T635 | 1666-1669 | DT | denotes | The |
| T636 | 1670-1680 | JJ | denotes | N-terminal |
| T637 | 1681-1691 | JJ | denotes | regulatory |
| T638 | 1692-1698 | NN | denotes | region |
| T639 | 1699-1701 | IN | denotes | of |
| T640 | 1702-1705 | NN | denotes | PKD |
| T641 | 1706-1713 | NN | denotes | enzymes |
| T642 | 1714-1722 | VB | denotes | contains |
| T643 | 1723-1724 | DT | denotes | a |
| T644 | 1725-1728 | NN | denotes | DAG |
| T645 | 1729-1736 | NN | denotes | binding |
| T646 | 1737-1743 | NN | denotes | domain |
| T647 | 1744-1747 | CC | denotes | and |
| T648 | 1748-1754 | JJ | denotes | direct |
| T649 | 1755-1762 | NN | denotes | binding |
| T650 | 1763-1765 | IN | denotes | of |
| T651 | 1766-1769 | NN | denotes | DAG |
| T652 | 1770-1774 | RB | denotes | also |
| T653 | 1775-1786 | VB | denotes | contributes |
| T654 | 1787-1789 | TO | denotes | to |
| T655 | 1790-1794 | NN | denotes | PKD1 |
| T656 | 1795-1805 | NN | denotes | activation |
| T657 | 1806-1807 | -LRB- | denotes | [ |
| T658 | 1807-1808 | CD | denotes | 7 |
| T659 | 1808-1809 | -RRB- | denotes | ] |
| T660 | 1810-1812 | RB | denotes | as |
| T661 | 1813-1817 | RB | denotes | well |
| T662 | 1818-1820 | IN | denotes | as |
| T663 | 1821-1831 | VB | denotes | regulating |
| T664 | 1832-1835 | DT | denotes | the |
| T665 | 1836-1843 | JJ | denotes | spatial |
| T666 | 1844-1852 | NN | denotes | location |
| T667 | 1853-1855 | IN | denotes | of |
| T668 | 1856-1859 | NN | denotes | PKD |
| T669 | 1860-1867 | NN | denotes | enzymes |
| T670 | 1868-1874 | IN | denotes | within |
| T671 | 1875-1880 | NN | denotes | cells |
| T672 | 1881-1882 | -LRB- | denotes | [ |
| T673 | 1882-1886 | CD | denotes | 8–12 |
| T674 | 1886-1887 | -RRB- | denotes | ] |
| T675 | 1889-1892 | NN | denotes | PKD |
| T676 | 1893-1900 | NN | denotes | enzymes |
| T677 | 1901-1905 | VB | denotes | have |
| T678 | 1906-1910 | VB | denotes | been |
| T679 | 1911-1919 | VB | denotes | proposed |
| T680 | 1920-1922 | TO | denotes | to |
| T681 | 1923-1931 | VB | denotes | regulate |
| T682 | 1932-1940 | JJ | denotes | numerous |
| T683 | 1941-1949 | JJ | denotes | cellular |
| T684 | 1950-1959 | NN | denotes | functions |
| T685 | 1959-1960 | -COMMA- | denotes | , |
| T686 | 1961-1970 | VB | denotes | including |
| T687 | 1971-1975 | NN | denotes | cell |
| T688 | 1976-1989 | NN | denotes | proliferation |
| T689 | 1990-1991 | -LRB- | denotes | [ |
| T690 | 1991-1996 | CD | denotes | 13–16 |
| T691 | 1996-1997 | -RRB- | denotes | ] |
| T692 | 1997-1998 | -COMMA- | denotes | , |
| T693 | 1999-2013 | JJ | denotes | anti-apoptotic |
| T694 | 2014-2021 | NN | denotes | signals |
| T695 | 2022-2023 | -LRB- | denotes | [ |
| T696 | 2023-2028 | CD | denotes | 17,18 |
| T697 | 2028-2029 | -RRB- | denotes | ] |
| T698 | 2030-2033 | CC | denotes | and |
| T699 | 2034-2043 | NN | denotes | thymocyte |
| T700 | 2044-2055 | NN | denotes | development |
| T701 | 2056-2057 | -LRB- | denotes | [ |
| T702 | 2057-2059 | CD | denotes | 19 |
| T703 | 2059-2060 | -RRB- | denotes | ] |
| T704 | 2062-2072 | NN | denotes | Expression |
| T705 | 2073-2075 | IN | denotes | of |
| T706 | 2076-2082 | JJ | denotes | mutant |
| T707 | 2083-2096 | RB | denotes | catalytically |
| T708 | 2097-2105 | JJ | denotes | inactive |
| T709 | 2106-2109 | CC | denotes | and |
| T710 | 2110-2124 | RB | denotes | constitutively |
| T711 | 2125-2134 | VB | denotes | activated |
| T712 | 2135-2139 | NN | denotes | PKDs |
| T713 | 2140-2143 | MD | denotes | can |
| T714 | 2144-2148 | RB | denotes | also |
| T715 | 2149-2155 | VB | denotes | modify |
| T716 | 2156-2161 | NNP | denotes | Golgi |
| T717 | 2162-2170 | NN | denotes | function |
| T718 | 2170-2171 | -COMMA- | denotes | , |
| T719 | 2172-2176 | NN | denotes | cell |
| T720 | 2177-2185 | NN | denotes | adhesion |
| T721 | 2186-2189 | CC | denotes | and |
| T722 | 2190-2194 | NN | denotes | cell |
| T723 | 2195-2203 | NN | denotes | motility |
| T724 | 2204-2205 | -LRB- | denotes | ( |
| T725 | 2205-2213 | VB | denotes | reviewed |
| T726 | 2214-2216 | IN | denotes | in |
| T727 | 2217-2218 | -LRB- | denotes | [ |
| T728 | 2218-2220 | CD | denotes | 20 |
| T729 | 2220-2221 | -RRB- | denotes | ] |
| T730 | 2221-2222 | -RRB- | denotes | ) |
| T731 | 2224-2226 | IN | denotes | In |
| T732 | 2227-2237 | JJ | denotes | particular |
| T733 | 2237-2238 | -COMMA- | denotes | , |
| T734 | 2239-2243 | NN | denotes | PKDs |
| T735 | 2244-2248 | VB | denotes | have |
| T736 | 2249-2253 | VB | denotes | been |
| T737 | 2254-2260 | RB | denotes | widely |
| T738 | 2261-2267 | VB | denotes | linked |
| T739 | 2268-2270 | TO | denotes | to |
| T740 | 2271-2274 | DT | denotes | the |
| T741 | 2275-2285 | NN | denotes | activation |
| T742 | 2286-2288 | IN | denotes | of |
| T743 | 2289-2292 | DT | denotes | the |
| T744 | 2293-2297 | NN | denotes | NFκB |
| T745 | 2298-2311 | NN | denotes | transcription |
| T746 | 2312-2318 | NN | denotes | factor |
| T747 | 2319-2322 | CC | denotes | and |
| T748 | 2323-2325 | IN | denotes | in |
| T749 | 2326-2336 | VB | denotes | regulating |
| T750 | 2337-2341 | NN | denotes | cell |
| T751 | 2342-2350 | NN | denotes | survival |
| T752 | 2351-2357 | IN | denotes | during |
| T753 | 2358-2367 | JJ | denotes | oxidative |
| T754 | 2368-2374 | NN | denotes | stress |
| T755 | 2375-2376 | -LRB- | denotes | [ |
| T756 | 2376-2384 | CD | denotes | 17,21–23 |
| T757 | 2384-2385 | -RRB- | denotes | ] |
| T758 | 2387-2394 | DT | denotes | Another |
| T759 | 2395-2403 | RB | denotes | recently |
| T760 | 2404-2412 | VB | denotes | proposed |
| T761 | 2413-2417 | NN | denotes | PKD1 |
| T762 | 2418-2427 | NN | denotes | substrate |
| T763 | 2428-2430 | VB | denotes | is |
| T764 | 2431-2436 | NN | denotes | HSP27 |
| T765 | 2437-2438 | -LRB- | denotes | [ |
| T766 | 2438-2440 | CD | denotes | 24 |
| T767 | 2440-2441 | -RRB- | denotes | ] |
| T768 | 2441-2442 | -COMMA- | denotes | , |
| T769 | 2443-2444 | DT | denotes | a |
| T770 | 2445-2450 | JJ | denotes | small |
| T771 | 2451-2455 | NN | denotes | heat |
| T772 | 2456-2461 | NN | denotes | shock |
| T773 | 2462-2469 | NN | denotes | protein |
| T774 | 2470-2478 | VB | denotes | involved |
| T775 | 2479-2481 | IN | denotes | in |
| T776 | 2482-2492 | VB | denotes | regulating |
| T777 | 2493-2497 | NN | denotes | cell |
| T778 | 2498-2507 | NN | denotes | migration |
| T779 | 2508-2511 | CC | denotes | and |
| T780 | 2512-2516 | NN | denotes | cell |
| T781 | 2517-2525 | NN | denotes | survival |
| T782 | 2526-2527 | -LRB- | denotes | [ |
| T783 | 2527-2529 | CD | denotes | 25 |
| T784 | 2529-2530 | -RRB- | denotes | ] |
| T785 | 2532-2534 | DT | denotes | An |
| T786 | 2535-2544 | JJ | denotes | essential |
| T787 | 2545-2549 | NN | denotes | role |
| T788 | 2550-2553 | IN | denotes | for |
| T789 | 2554-2557 | NN | denotes | PKD |
| T790 | 2558-2565 | NN | denotes | enzymes |
| T791 | 2566-2568 | IN | denotes | in |
| T792 | 2569-2579 | VB | denotes | regulating |
| T793 | 2580-2585 | NN | denotes | class |
| T794 | 2586-2588 | CD | denotes | II |
| T795 | 2589-2596 | NN | denotes | histone |
| T796 | 2597-2609 | NN | denotes | deacetylases |
| T797 | 2610-2611 | -LRB- | denotes | ( |
| T798 | 2611-2616 | NN | denotes | HDACs |
| T799 | 2616-2617 | -RRB- | denotes | ) |
| T800 | 2617-2618 | -COMMA- | denotes | , |
| T801 | 2619-2626 | NN | denotes | enzymes |
| T802 | 2627-2631 | WDT | denotes | that |
| T803 | 2632-2639 | VB | denotes | repress |
| T804 | 2640-2654 | JJ | denotes | MEF2-dependent |
| T805 | 2655-2659 | NN | denotes | gene |
| T806 | 2660-2673 | NN | denotes | transcription |
| T807 | 2673-2674 | -COMMA- | denotes | , |
| T808 | 2675-2678 | VB | denotes | has |
| T809 | 2679-2683 | RB | denotes | also |
| T810 | 2684-2688 | VB | denotes | been |
| T811 | 2689-2701 | VB | denotes | demonstrated |
| T812 | 2702-2703 | -LRB- | denotes | [ |
| T813 | 2703-2710 | CD | denotes | 1,26–28 |
| T814 | 2710-2711 | -RRB- | denotes | ] |
| T815 | 2713-2715 | TO | denotes | To |
| T816 | 2716-2727 | VB | denotes | investigate |
| T817 | 2728-2731 | DT | denotes | the |
| T818 | 2732-2742 | JJ | denotes | biological |
| T819 | 2743-2747 | NN | denotes | role |
| T820 | 2748-2750 | IN | denotes | of |
| T821 | 2751-2755 | NN | denotes | PKDs |
| T822 | 2756-2758 | PRP | denotes | we |
| T823 | 2759-2763 | VB | denotes | have |
| T824 | 2764-2773 | VB | denotes | generated |
| T825 | 2774-2778 | NN | denotes | DT40 |
| T826 | 2779-2780 | NN | denotes | B |
| T827 | 2781-2785 | NN | denotes | cell |
| T828 | 2786-2791 | NN | denotes | lines |
| T829 | 2792-2796 | WDT | denotes | that |
| T830 | 2797-2801 | VB | denotes | lack |
| T831 | 2802-2812 | NN | denotes | expression |
| T832 | 2813-2815 | IN | denotes | of |
| T833 | 2816-2819 | CD | denotes | one |
| T834 | 2820-2822 | CC | denotes | or |
| T835 | 2823-2827 | JJ | denotes | more |
| T836 | 2828-2835 | NN | denotes | members |
| T837 | 2836-2838 | IN | denotes | of |
| T838 | 2839-2842 | DT | denotes | the |
| T839 | 2843-2846 | NN | denotes | PKD |
| T840 | 2847-2853 | NN | denotes | family |
| T841 | 2854-2855 | -LRB- | denotes | [ |
| T842 | 2855-2856 | CD | denotes | 1 |
| T843 | 2856-2857 | -RRB- | denotes | ] |
| T844 | 2857-2858 | -COMMA- | denotes | , |
| T845 | 2859-2867 | VB | denotes | allowing |
| T846 | 2868-2870 | PRP | denotes | us |
| T847 | 2871-2873 | TO | denotes | to |
| T848 | 2874-2885 | VB | denotes | investigate |
| T849 | 2886-2889 | DT | denotes | the |
| T850 | 2890-2898 | NN | denotes | function |
| T851 | 2898-2899 | -LRB- | denotes | ( |
| T852 | 2899-2900 | NN | denotes | s |
| T853 | 2900-2901 | -RRB- | denotes | ) |
| T854 | 2902-2904 | IN | denotes | of |
| T855 | 2905-2908 | NN | denotes | PKD |
| T856 | 2909-2917 | NN | denotes | isoforms |
| T857 | 2918-2927 | VB | denotes | following |
| T858 | 2928-2929 | NN | denotes | B |
| T859 | 2930-2934 | NN | denotes | cell |
| T860 | 2935-2942 | NN | denotes | antigen |
| T861 | 2943-2951 | NN | denotes | receptor |
| T862 | 2952-2953 | -LRB- | denotes | ( |
| T863 | 2953-2956 | NN | denotes | BCR |
| T864 | 2956-2957 | -RRB- | denotes | ) |
| T865 | 2958-2969 | NN | denotes | stimulation |
| T866 | 2969-2970 | -COMMA- | denotes | , |
| T867 | 2971-2973 | RB | denotes | as |
| T868 | 2974-2978 | RB | denotes | well |
| T869 | 2979-2989 | VB | denotes | addressing |
| T870 | 2990-2993 | DT | denotes | the |
| T871 | 2994-2999 | NN | denotes | issue |
| T872 | 3000-3002 | IN | denotes | of |
| T873 | 3003-3013 | JJ | denotes | functional |
| T874 | 3014-3024 | NN | denotes | redundancy |
| T875 | 3025-3032 | IN | denotes | between |
| T876 | 3033-3036 | DT | denotes | the |
| T877 | 3037-3046 | JJ | denotes | different |
| T878 | 3047-3050 | NN | denotes | PKD |
| T879 | 3051-3057 | NN | denotes | family |
| T880 | 3058-3065 | NN | denotes | members |
| T881 | 3067-3075 | JJ | denotes | Previous |
| T882 | 3076-3083 | NN | denotes | studies |
| T883 | 3084-3088 | VB | denotes | have |
| T884 | 3089-3094 | VB | denotes | shown |
| T885 | 3095-3099 | IN | denotes | that |
| T886 | 3100-3104 | NN | denotes | PKDs |
| T887 | 3105-3108 | VB | denotes | are |
| T888 | 3109-3122 | JJ | denotes | indispensable |
| T889 | 3123-3126 | IN | denotes | for |
| T890 | 3127-3131 | NN | denotes | HDAC |
| T891 | 3132-3142 | NN | denotes | regulation |
| T892 | 3143-3145 | IN | denotes | in |
| T893 | 3146-3147 | NN | denotes | B |
| T894 | 3148-3153 | NN | denotes | cells |
| T895 | 3154-3155 | -LRB- | denotes | [ |
| T896 | 3155-3156 | CD | denotes | 1 |
| T897 | 3156-3157 | -RRB- | denotes | ] |
| T898 | 3159-3165 | RB | denotes | Herein |
| T899 | 3166-3168 | PRP | denotes | we |
| T900 | 3169-3173 | VB | denotes | show |
| T901 | 3174-3178 | IN | denotes | that |
| T902 | 3179-3183 | NN | denotes | PKDs |
| T903 | 3184-3187 | VB | denotes | are |
| T904 | 3188-3192 | RB | denotes | also |
| T905 | 3193-3206 | JJ | denotes | indispensable |
| T906 | 3207-3210 | IN | denotes | for |
| T907 | 3211-3216 | NN | denotes | HSP27 |
| T908 | 3217-3232 | NN | denotes | phosphorylation |
| T909 | 3233-3235 | IN | denotes | in |
| T910 | 3236-3237 | NN | denotes | B |
| T911 | 3238-3243 | NN | denotes | cells |
| T912 | 3245-3252 | RB | denotes | However |
| T913 | 3252-3253 | -COMMA- | denotes | , |
| T914 | 3254-3262 | JJ | denotes | PKD-null |
| T915 | 3263-3267 | NN | denotes | DT40 |
| T916 | 3268-3269 | NN | denotes | B |
| T917 | 3270-3275 | NN | denotes | cells |
| T918 | 3276-3279 | VB | denotes | are |
| T919 | 3280-3286 | JJ | denotes | viable |
| T920 | 3287-3290 | CC | denotes | and |
| T921 | 3291-3302 | VB | denotes | proliferate |
| T922 | 3303-3311 | RB | denotes | normally |
| T923 | 3313-3321 | RB | denotes | Moreover |
| T924 | 3321-3322 | -COMMA- | denotes | , |
| T925 | 3323-3327 | NN | denotes | loss |
| T926 | 3328-3330 | IN | denotes | of |
| T927 | 3331-3334 | DT | denotes | the |
| T928 | 3335-3341 | JJ | denotes | entire |
| T929 | 3342-3350 | JJ | denotes | cellular |
| T930 | 3351-3355 | NN | denotes | pool |
| T931 | 3356-3358 | IN | denotes | of |
| T932 | 3359-3362 | NN | denotes | PKD |
| T933 | 3363-3367 | VB | denotes | does |
| T934 | 3368-3371 | RB | denotes | not |
| T935 | 3372-3382 | RB | denotes | critically |
| T936 | 3383-3389 | VB | denotes | affect |
| T937 | 3390-3399 | JJ | denotes | oxidative |
| T938 | 3400-3406 | NN | denotes | stress |
| T939 | 3407-3416 | NN | denotes | responses |
| T940 | 3417-3419 | IN | denotes | in |
| T941 | 3420-3421 | NN | denotes | B |
| T942 | 3422-3427 | NN | denotes | cells |
| T943 | 3428-3431 | CC | denotes | nor |
| T944 | 3432-3434 | VB | denotes | do |
| T945 | 3435-3438 | NN | denotes | PKD |
| T946 | 3439-3446 | NN | denotes | kinases |
| T947 | 3447-3451 | VB | denotes | play |
| T948 | 3452-3454 | DT | denotes | an |
| T949 | 3455-3464 | JJ | denotes | essential |
| T950 | 3465-3469 | NN | denotes | role |
| T951 | 3470-3472 | IN | denotes | in |
| T952 | 3473-3483 | VB | denotes | regulating |
| T953 | 3484-3488 | NN | denotes | NFκB |
| T954 | 3489-3504 | JJ | denotes | transcriptional |
| T955 | 3505-3513 | NN | denotes | activity |
| T956 | 3515-3523 | RB | denotes | Together |
| T957 | 3523-3524 | -COMMA- | denotes | , |
| T958 | 3525-3530 | DT | denotes | these |
| T959 | 3531-3539 | NN | denotes | findings |
| T960 | 3540-3546 | VB | denotes | reveal |
| T961 | 3547-3551 | IN | denotes | that |
| T962 | 3552-3554 | IN | denotes | in |
| T963 | 3555-3556 | NN | denotes | B |
| T964 | 3557-3568 | NN | denotes | lymphocytes |
| T965 | 3568-3569 | -COMMA- | denotes | , |
| T966 | 3570-3573 | NN | denotes | PKD |
| T967 | 3574-3581 | NN | denotes | kinases |
| T968 | 3582-3585 | VB | denotes | are |
| T969 | 3586-3589 | RB | denotes | not |
| T970 | 3590-3598 | JJ | denotes | critical |
| T971 | 3599-3609 | NN | denotes | regulators |
| T972 | 3610-3612 | IN | denotes | of |
| T973 | 3613-3617 | JJ | denotes | many |
| T974 | 3618-3620 | IN | denotes | of |
| T975 | 3621-3624 | DT | denotes | the |
| T976 | 3625-3633 | JJ | denotes | cellular |
| T977 | 3634-3643 | NN | denotes | processes |
| T978 | 3644-3654 | RB | denotes | previously |
| T979 | 3655-3663 | VB | denotes | ascribed |
| T980 | 3664-3666 | TO | denotes | to |
| T981 | 3667-3671 | PRP | denotes | them |
| T982 | 3672-3674 | IN | denotes | in |
| T983 | 3675-3680 | JJ | denotes | other |
| T984 | 3681-3689 | JJ | denotes | cellular |
| T985 | 3690-3697 | NN | denotes | systems |
| T1674 | 0-3735 | NN | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell |
| T1675 | 3736-3743 | NN | denotes | culture |
| T1676 | 3743-3744 | -COMMA- | denotes | , |
| T1677 | 3745-3754 | JJ | denotes | transient |
| T1678 | 3755-3768 | NN | denotes | transfections |
| T1679 | 3769-3772 | CC | denotes | and |
| T1680 | 3773-3777 | NN | denotes | cell |
| T1681 | 3778-3789 | NN | denotes | stimulation |
| T1682 | 3790-3793 | DT | denotes | The |
| T1683 | 3794-3804 | NN | denotes | generation |
| T1684 | 3804-3805 | -COMMA- | denotes | , |
| T1685 | 3806-3813 | NN | denotes | culture |
| T1686 | 3814-3817 | CC | denotes | and |
| T1687 | 3818-3828 | NN | denotes | activation |
| T1688 | 3829-3831 | IN | denotes | of |
| T1689 | 3832-3839 | NN | denotes | PKD1−/− |
| T1690 | 3839-3840 | -COMMA- | denotes | , |
| T1691 | 3841-3848 | NN | denotes | PKD3−/− |
| T1692 | 3849-3852 | CC | denotes | and |
| T1693 | 3853-3862 | NN | denotes | PKD1/3−/− |
| T1694 | 3863-3871 | NN | denotes | knockout |
| T1695 | 3872-3876 | NN | denotes | DT40 |
| T1696 | 3877-3878 | NN | denotes | B |
| T1697 | 3879-3883 | NN | denotes | cell |
| T1698 | 3884-3889 | NN | denotes | lines |
| T1699 | 3890-3894 | VB | denotes | have |
| T1700 | 3895-3899 | VB | denotes | been |
| T1701 | 3900-3909 | VB | denotes | described |
| T1702 | 3910-3920 | RB | denotes | previously |
| T1703 | 3921-3922 | -LRB- | denotes | [ |
| T1704 | 3922-3923 | CD | denotes | 1 |
| T1705 | 3923-3924 | -RRB- | denotes | ] |
| T1706 | 3926-3931 | NN | denotes | Cells |
| T1707 | 3932-3936 | VB | denotes | were |
| T1708 | 3937-3942 | VB | denotes | lysed |
| T1709 | 3943-3946 | CC | denotes | and |
| T1710 | 3947-3954 | NN | denotes | protein |
| T1711 | 3955-3963 | NN | denotes | extracts |
| T1712 | 3964-3968 | VB | denotes | were |
| T1713 | 3969-3977 | VB | denotes | analysed |
| T1714 | 3978-3980 | IN | denotes | in |
| T1715 | 3981-3988 | JJ | denotes | Western |
| T1716 | 3989-3997 | NN | denotes | blotting |
| T1717 | 3998-4009 | NN | denotes | experiments |
| T1718 | 4010-4012 | IN | denotes | as |
| T1719 | 4013-4023 | RB | denotes | previously |
| T1720 | 4024-4033 | VB | denotes | described |
| T1721 | 4034-4035 | -LRB- | denotes | [ |
| T26 | 0-7 | NN | denotes | Protein |
| T27 | 8-14 | NN | denotes | kinase |
| T28 | 15-16 | NN | denotes | D |
| T29 | 17-24 | NN | denotes | enzymes |
| T30 | 25-28 | VB | denotes | are |
| T31 | 29-40 | JJ | denotes | dispensable |
| T32 | 41-44 | IN | denotes | for |
| T33 | 45-58 | NN | denotes | proliferation |
| T34 | 58-59 | -COMMA- | denotes | , |
| T35 | 60-68 | NN | denotes | survival |
| T36 | 69-72 | CC | denotes | and |
| T37 | 73-80 | NN | denotes | antigen |
| T38 | 81-99 | JJ | denotes | receptor-regulated |
| T39 | 100-104 | NN | denotes | NFκB |
| T40 | 105-113 | NN | denotes | activity |
| T41 | 114-116 | IN | denotes | in |
| T42 | 117-127 | NN | denotes | vertebrate |
| T43 | 128-135 | NN | denotes | B-cells |
| T44 | 147-149 | TO | denotes | To |
| T45 | 150-161 | VB | denotes | investigate |
| T46 | 162-165 | DT | denotes | the |
| T47 | 166-176 | NN | denotes | importance |
| T48 | 177-179 | IN | denotes | of |
| T1722 | 4035-4036 | CD | denotes | 1 |
| T1723 | 4036-4037 | -RRB- | denotes | ] |
| T1724 | 4039-4054 | NN | denotes | Chloramphenicol |
| T1725 | 4055-4061 | NN | denotes | acetyl |
| T1726 | 4062-4073 | NN | denotes | transferase |
| T1727 | 4074-4080 | NN | denotes | assays |
| T1728 | 4081-4085 | VB | denotes | have |
| T1729 | 4086-4090 | VB | denotes | been |
| T1730 | 4091-4100 | VB | denotes | described |
| T1731 | 4101-4111 | RB | denotes | previously |
| T1732 | 4112-4113 | -LRB- | denotes | [ |
| T1733 | 4113-4115 | CD | denotes | 29 |
| T1734 | 4115-4116 | -RRB- | denotes | ] |
| T1868 | 0-4128 | NN | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell culture, transient transfections and cell stimulation The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. Chloramphenicol acetyl transferase assays have been described previously [29]. 2.2 sIgM |
| T1869 | 4129-4137 | NN | denotes | staining |
| R9 | T51 | T52 | arg1Of | D,( |
| R10 | T53 | T52 | arg2Of | PKD,( |
| R11 | T54 | T52 | arg3Of | ),( |
| R15 | T56 | T57 | arg1Of | we,generated |
| R17 | T63 | T57 | arg2Of | line,generated |
| R18 | T63 | T58 | arg1Of | line,a |
| R19 | T63 | T59 | arg1Of | line,PKD-null |
| R20 | T63 | T60 | arg1Of | line,DT40 |
| R21 | T63 | T61 | arg1Of | line,B-lymphocyte |
| R22 | T63 | T62 | arg1Of | line,cell |
| R23 | T65 | T66 | arg1Of | we,have |
| R24 | T65 | T67 | arg1Of | we,shown |
| R25 | T67 | T64 | arg1Of | shown,Previously |
| R26 | T67 | T66 | arg2Of | shown,have |
| R27 | T69 | T70 | arg1Of | PKDs,have |
| R28 | T70 | T67 | arg2Of | have,shown |
| R29 | T70 | T68 | arg1Of | have,that |
| R30 | T70 | T74 | arg1Of | have,in |
| R31 | T73 | T70 | arg2Of | role,have |
| R32 | T73 | T71 | arg1Of | role,an |
| R33 | T73 | T72 | arg1Of | role,essential |
| R34 | T75 | T74 | arg2Of | regulating,in |
| R35 | T75 | T80 | arg1Of | regulating,in |
| R36 | T79 | T75 | arg2Of | deacetylases,regulating |
| R37 | T79 | T76 | arg1Of | deacetylases,class |
| R38 | T79 | T77 | arg1Of | deacetylases,II |
| R39 | T79 | T78 | arg1Of | deacetylases,histone |
| R40 | T82 | T80 | arg2Of | B-cells,in |
| R41 | T82 | T81 | arg1Of | B-cells,DT40 |
| R42 | T84 | T85 | arg1Of | Matthews,"," |
| R43 | T85 | T87 | arg1Of | ",","," |
| R44 | T86 | T85 | arg2Of | S.A.,"," |
| R45 | T87 | T89 | arg1Of | ",","," |
| R46 | T88 | T87 | arg2Of | Liu,"," |
| R47 | T89 | T91 | arg1Of | ",","," |
| R48 | T90 | T89 | arg2Of | P.,"," |
| R49 | T91 | T93 | arg1Of | ",","," |
| R50 | T92 | T91 | arg2Of | Spitaler,"," |
| R51 | T93 | T95 | arg1Of | ",","," |
| R52 | T94 | T93 | arg2Of | M.,"," |
| R53 | T95 | T105 | arg1Of | ",","," |
| R54 | T96 | T97 | arg1Of | Olson,"," |
| R55 | T97 | T99 | arg1Of | ",","," |
| R56 | T98 | T97 | arg2Of | E.N.,"," |
| R57 | T99 | T101 | arg1Of | ",","," |
| R58 | T100 | T99 | arg2Of | McKinsey,"," |
| R59 | T101 | T103 | arg1Of | ",","," |
| R60 | T102 | T101 | arg2Of | T.A.,"," |
| R61 | T103 | T95 | arg2Of | ",","," |
| R62 | T104 | T103 | arg2Of | Cantrell,"," |
| R63 | T105 | T107 | arg1Of | ",",and |
| R64 | T106 | T105 | arg2Of | D.A.,"," |
| R65 | T108 | T109 | arg1Of | Scharenberg,"," |
| R66 | T109 | T107 | arg2Of | ",",and |
| R67 | T110 | T109 | arg2Of | A.M.,"," |
| R68 | T112 | T111 | arg2Of | 2006,( |
| R69 | T113 | T111 | arg3Of | ),( |
| R70 | T115 | T80 | arg3Of | role,in |
| R71 | T115 | T83 | arg1Of | role,[ |
| R72 | T115 | T84 | arg1Of | role,Matthews |
| R73 | T115 | T86 | arg1Of | role,S.A. |
| R74 | T115 | T88 | arg1Of | role,Liu |
| R75 | T115 | T90 | arg1Of | role,P. |
| R76 | T115 | T92 | arg1Of | role,Spitaler |
| R77 | T115 | T94 | arg1Of | role,M. |
| R78 | T115 | T96 | arg1Of | role,Olson |
| R79 | T115 | T98 | arg1Of | role,E.N. |
| R80 | T115 | T100 | arg1Of | role,McKinsey |
| R81 | T115 | T102 | arg1Of | role,T.A. |
| R82 | T115 | T104 | arg1Of | role,Cantrell |
| R83 | T115 | T106 | arg1Of | role,D.A. |
| R84 | T115 | T108 | arg1Of | role,Scharenberg |
| R85 | T115 | T110 | arg1Of | role,A.M. |
| R86 | T115 | T111 | arg1Of | role,( |
| R87 | T115 | T114 | arg1Of | role,Essential |
| R88 | T115 | T116 | arg1Of | role,for |
| R89 | T115 | T122 | arg1Of | role,in |
| R90 | T121 | T116 | arg2Of | kinases,for |
| R91 | T121 | T117 | arg1Of | kinases,protein |
| R92 | T121 | T118 | arg1Of | kinases,kinase |
| R93 | T121 | T119 | arg1Of | kinases,D |
| R94 | T121 | T120 | arg1Of | kinases,family |
| R95 | T124 | T122 | arg2Of | regulation,in |
| R96 | T124 | T123 | arg1Of | regulation,the |
| R97 | T124 | T125 | arg1Of | regulation,of |
| R98 | T129 | T125 | arg2Of | deacetylases,of |
| R99 | T129 | T126 | arg1Of | deacetylases,class |
| R100 | T129 | T127 | arg1Of | deacetylases,II |
| R101 | T129 | T128 | arg1Of | deacetylases,histone |
| R102 | T129 | T130 | arg1Of | deacetylases,in |
| R103 | T132 | T130 | arg2Of | lymphocytes,in |
| R104 | T132 | T131 | arg1Of | lymphocytes,B |
| R105 | T135 | T133 | arg1Of | Biol.,Mol. |
| R106 | T135 | T134 | arg1Of | Biol.,Cell |
| R107 | T135 | T136 | arg1Of | Biol.,26 |
| R108 | T135 | T137 | arg1Of | Biol.,"," |
| R109 | T138 | T137 | arg2Of | 1569–1577,"," |
| R110 | T138 | T139 | arg1Of | 1569–1577,] |
| R111 | T140 | T142 | arg1Of | We,show |
| R112 | T142 | T141 | arg1Of | show,now |
| R113 | T144 | T145 | arg1Of | PKDs,are |
| R114 | T144 | T147 | arg2Of | PKDs,required |
| R115 | T144 | T149 | arg1Of | PKDs,regulate |
| R116 | T147 | T142 | arg2Of | required,show |
| R117 | T147 | T143 | arg1Of | required,that |
| R118 | T147 | T145 | arg2Of | required,are |
| R119 | T147 | T146 | arg1Of | required,also |
| R120 | T149 | T147 | arg3Of | regulate,required |
| R121 | T149 | T148 | arg1Of | regulate,to |
| R122 | T149 | T152 | arg1Of | regulate,in |
| R123 | T151 | T149 | arg2Of | phosphorylation,regulate |
| R124 | T151 | T150 | arg1Of | phosphorylation,HSP27 |
| R125 | T154 | T152 | arg2Of | B-cells,in |
| R126 | T154 | T153 | arg1Of | B-cells,DT40 |
| R127 | T158 | T157 | arg2Of | contrast,in |
| R128 | T158 | T159 | arg1Of | contrast,to |
| R129 | T161 | T159 | arg2Of | observations,to |
| R130 | T161 | T160 | arg1Of | observations,previous |
| R131 | T161 | T162 | arg1Of | observations,in |
| R132 | T165 | T162 | arg2Of | types,in |
| R133 | T165 | T163 | arg1Of | types,other |
| R134 | T165 | T164 | arg1Of | types,cell |
| R135 | T168 | T167 | arg1Of | enzymes,PKD |
| R136 | T168 | T169 | arg1Of | enzymes,do |
| R137 | T168 | T171 | arg1Of | enzymes,regulate |
| R138 | T171 | T155 | arg1Of | regulate,However |
| R139 | T171 | T156 | arg1Of | regulate,"," |
| R140 | T171 | T157 | arg1Of | regulate,in |
| R141 | T171 | T166 | arg1Of | regulate,"," |
| R142 | T171 | T169 | arg2Of | regulate,do |
| R143 | T171 | T170 | arg1Of | regulate,not |
| R144 | T174 | T171 | arg2Of | processes,regulate |
| R145 | T174 | T172 | arg1Of | processes,basic |
| R146 | T174 | T173 | arg1Of | processes,cellular |
| R147 | T174 | T176 | arg1Of | processes,as |
| R148 | T176 | T175 | arg1Of | as,such |
| R149 | T177 | T178 | arg1Of | proliferation,or |
| R150 | T178 | T182 | arg1Of | or,nor |
| R151 | T180 | T178 | arg2Of | responses,or |
| R152 | T180 | T179 | arg1Of | responses,survival |
| R153 | T182 | T176 | arg2Of | nor,as |
| R154 | T182 | T181 | arg1Of | nor,"," |
| R155 | T185 | T182 | arg2Of | activity,nor |
| R156 | T185 | T183 | arg1Of | activity,NFκB |
| R157 | T185 | T184 | arg1Of | activity,transcriptional |
| R158 | T185 | T186 | arg1Of | activity,downstream |
| R159 | T186 | T187 | arg1Of | downstream,of |
| R160 | T192 | T187 | arg2Of | receptor,of |
| R161 | T192 | T188 | arg1Of | receptor,the |
| R162 | T192 | T189 | arg1Of | receptor,B |
| R163 | T192 | T190 | arg1Of | receptor,cell |
| R164 | T192 | T191 | arg1Of | receptor,antigen |
| R165 | T195 | T196 | arg1Of | PKDs,have |
| R166 | T196 | T193 | arg1Of | have,Thus |
| R167 | T196 | T194 | arg1Of | have,"," |
| R168 | T196 | T200 | arg1Of | have,in |
| R169 | T199 | T196 | arg2Of | role,have |
| R170 | T199 | T197 | arg1Of | role,a |
| R171 | T199 | T198 | arg1Of | role,selective |
| R172 | T203 | T200 | arg2Of | biology,in |
| R173 | T203 | T201 | arg1Of | biology,DT40 |
| R174 | T203 | T202 | arg1Of | biology,B-cell |
| R430 | T535 | T532 | arg1Of | D,"1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The" |
| R431 | T535 | T533 | arg1Of | D,protein |
| R432 | T535 | T534 | arg1Of | D,kinase |
| R433 | T535 | T536 | arg1Of | D,( |
| R434 | T537 | T536 | arg2Of | PKD,( |
| R435 | T538 | T536 | arg3Of | ),( |
| R436 | T541 | T535 | arg1Of | family,D |
| R437 | T541 | T539 | arg1Of | family,serine/threonine |
| R438 | T541 | T540 | arg1Of | family,kinase |
| R439 | T541 | T542 | arg1Of | family,has |
| R440 | T544 | T542 | arg2Of | members,has |
| R441 | T544 | T543 | arg1Of | members,three |
| R442 | T544 | T545 | arg1Of | members,: |
| R443 | T546 | T547 | arg1Of | PKD1,"," |
| R444 | T547 | T549 | arg1Of | ",",and |
| R445 | T548 | T547 | arg2Of | PKD2,"," |
| R446 | T549 | T545 | arg2Of | and,: |
| R447 | T550 | T549 | arg2Of | PKD3,and |
| R448 | T553 | T551 | arg1Of | types,Most |
| R449 | T553 | T552 | arg1Of | types,cell |
| R450 | T553 | T554 | arg1Of | types,express |
| R451 | T554 | T560 | arg1Of | express,but |
| R452 | T557 | T555 | arg1Of | two,at |
| R453 | T557 | T556 | arg1Of | two,least |
| R454 | T559 | T554 | arg2Of | isoforms,express |
| R455 | T559 | T557 | arg1Of | isoforms,two |
| R456 | T559 | T558 | arg1Of | isoforms,PKD |
| R457 | T562 | T561 | arg1Of | enzymes,PKD |
| R458 | T562 | T563 | arg1Of | enzymes,are |
| R459 | T562 | T566 | arg2Of | enzymes,expressed |
| R460 | T565 | T564 | arg1Of | highly,especially |
| R461 | T566 | T560 | arg2Of | expressed,but |
| R462 | T566 | T563 | arg2Of | expressed,are |
| R463 | T566 | T565 | arg1Of | expressed,highly |
| R464 | T566 | T567 | arg1Of | expressed,in |
| R465 | T569 | T567 | arg2Of | cells,in |
| R466 | T569 | T568 | arg1Of | cells,haematopoietic |
| R467 | T569 | T570 | arg1Of | cells,"," |
| R468 | T569 | T571 | arg1Of | cells,where |
| R469 | T572 | T573 | arg1Of | they,are |
| R470 | T572 | T574 | arg2Of | they,activated |
| R471 | T574 | T571 | arg2Of | activated,where |
| R472 | T574 | T573 | arg2Of | activated,are |
| R473 | T574 | T575 | arg1Of | activated,in |
| R474 | T574 | T581 | arg1Of | activated,[ |
| R475 | T575 | T577 | arg1Of | in,to |
| R476 | T576 | T575 | arg2Of | response,in |
| R477 | T580 | T575 | arg3Of | stimulation,in |
| R478 | T580 | T578 | arg1Of | stimulation,antigen |
| R479 | T580 | T579 | arg1Of | stimulation,receptors |
| R480 | T582 | T581 | arg2Of | "2,3",[ |
| R481 | T583 | T581 | arg3Of | ],[ |
| R482 | T587 | T584 | arg1Of | pathway,A |
| R483 | T587 | T585 | arg2Of | pathway,conserved |
| R484 | T587 | T586 | arg1Of | pathway,signalling |
| R485 | T587 | T588 | arg1Of | pathway,linking |
| R486 | T587 | T593 | arg1Of | pathway,involves |
| R487 | T590 | T588 | arg2Of | receptors,linking |
| R488 | T590 | T589 | arg1Of | receptors,antigen |
| R489 | T590 | T591 | arg1Of | receptors,to |
| R490 | T592 | T591 | arg2Of | PKDs,to |
| R491 | T595 | T594 | arg1Of | activation,the |
| R492 | T595 | T596 | arg1Of | activation,of |
| R493 | T595 | T598 | arg1Of | activation,and |
| R494 | T597 | T596 | arg2Of | PLCγ,of |
| R495 | T598 | T593 | arg2Of | and,involves |
| R496 | T601 | T598 | arg2Of | production,and |
| R497 | T601 | T599 | arg1Of | production,the |
| R498 | T601 | T600 | arg1Of | production,subsequent |
| R499 | T601 | T602 | arg1Of | production,of |
| R500 | T601 | T607 | arg1Of | production,which |
| R501 | T601 | T608 | arg1Of | production,stimulates |
| R502 | T603 | T602 | arg2Of | diacylglycerol,of |
| R503 | T603 | T604 | arg1Of | diacylglycerol,( |
| R504 | T605 | T604 | arg2Of | DAG,( |
| R505 | T606 | T604 | arg3Of | ),( |
| R506 | T609 | T610 | arg1Of | classical,and/or |
| R507 | T611 | T610 | arg2Of | novel,and/or |
| R508 | T614 | T608 | arg2Of | Cs,stimulates |
| R509 | T614 | T609 | arg1Of | Cs,classical |
| R510 | T614 | T611 | arg1Of | Cs,novel |
| R511 | T614 | T612 | arg1Of | Cs,protein |
| R512 | T614 | T613 | arg1Of | Cs,kinase |
| R513 | T614 | T615 | arg1Of | Cs,( |
| R514 | T614 | T618 | arg1Of | Cs,that |
| R515 | T614 | T619 | arg1Of | Cs,phosphorylate |
| R516 | T616 | T615 | arg2Of | PKC,( |
| R517 | T617 | T615 | arg3Of | ),( |
| R518 | T619 | T625 | arg1Of | phosphorylate,in |
| R519 | T624 | T619 | arg2Of | residues,phosphorylate |
| R520 | T624 | T620 | arg1Of | residues,two |
| R521 | T624 | T621 | arg1Of | residues,key |
| R522 | T624 | T622 | arg1Of | residues,regulatory |
| R523 | T624 | T623 | arg1Of | residues,serine |
| R524 | T628 | T625 | arg2Of | loop,in |
| R525 | T628 | T626 | arg1Of | loop,the |
| R526 | T628 | T627 | arg1Of | loop,activation |
| R527 | T628 | T629 | arg1Of | loop,of |
| R528 | T628 | T632 | arg1Of | loop,[ |
| R529 | T631 | T629 | arg2Of | kinases,of |
| R530 | T631 | T630 | arg1Of | kinases,PKD |
| R531 | T633 | T632 | arg2Of | 3–6,[ |
| R532 | T634 | T632 | arg3Of | ],[ |
| R533 | T638 | T635 | arg1Of | region,The |
| R534 | T638 | T636 | arg1Of | region,N-terminal |
| R535 | T638 | T637 | arg1Of | region,regulatory |
| R536 | T638 | T639 | arg1Of | region,of |
| R537 | T638 | T642 | arg1Of | region,contains |
| R538 | T641 | T639 | arg2Of | enzymes,of |
| R539 | T641 | T640 | arg1Of | enzymes,PKD |
| R540 | T642 | T647 | arg1Of | contains,and |
| R541 | T646 | T642 | arg2Of | domain,contains |
| R542 | T646 | T643 | arg1Of | domain,a |
| R543 | T646 | T644 | arg1Of | domain,DAG |
| R544 | T646 | T645 | arg1Of | domain,binding |
| R545 | T649 | T648 | arg1Of | binding,direct |
| R546 | T649 | T650 | arg1Of | binding,of |
| R547 | T649 | T653 | arg1Of | binding,contributes |
| R548 | T651 | T650 | arg2Of | DAG,of |
| R549 | T653 | T647 | arg2Of | contributes,and |
| R550 | T653 | T652 | arg1Of | contributes,also |
| R551 | T653 | T654 | arg1Of | contributes,to |
| R552 | T653 | T657 | arg1Of | contributes,[ |
| R553 | T653 | T661 | arg1Of | contributes,well |
| R554 | T653 | T662 | arg1Of | contributes,as |
| R555 | T656 | T654 | arg2Of | activation,to |
| R556 | T656 | T655 | arg1Of | activation,PKD1 |
| R557 | T658 | T657 | arg2Of | 7,[ |
| R558 | T659 | T657 | arg3Of | ],[ |
| R559 | T661 | T660 | arg1Of | well,as |
| R560 | T663 | T662 | arg2Of | regulating,as |
| R561 | T666 | T663 | arg2Of | location,regulating |
| R562 | T666 | T664 | arg1Of | location,the |
| R563 | T666 | T665 | arg1Of | location,spatial |
| R564 | T666 | T667 | arg1Of | location,of |
| R565 | T666 | T670 | arg1Of | location,within |
| R566 | T669 | T667 | arg2Of | enzymes,of |
| R567 | T669 | T668 | arg1Of | enzymes,PKD |
| R568 | T671 | T670 | arg2Of | cells,within |
| R569 | T671 | T672 | arg1Of | cells,[ |
| R570 | T673 | T672 | arg2Of | 8–12,[ |
| R571 | T674 | T672 | arg3Of | ],[ |
| R572 | T676 | T675 | arg1Of | enzymes,PKD |
| R573 | T676 | T677 | arg1Of | enzymes,have |
| R574 | T676 | T678 | arg1Of | enzymes,been |
| R575 | T676 | T679 | arg2Of | enzymes,proposed |
| R576 | T676 | T681 | arg1Of | enzymes,regulate |
| R577 | T679 | T677 | arg2Of | proposed,have |
| R578 | T679 | T678 | arg2Of | proposed,been |
| R579 | T681 | T679 | arg3Of | regulate,proposed |
| R580 | T681 | T680 | arg1Of | regulate,to |
| R581 | T684 | T681 | arg2Of | functions,regulate |
| R582 | T684 | T682 | arg1Of | functions,numerous |
| R583 | T684 | T683 | arg1Of | functions,cellular |
| R584 | T684 | T685 | arg1Of | functions,"," |
| R585 | T684 | T686 | arg1Of | functions,including |
| R586 | T688 | T687 | arg1Of | proliferation,cell |
| R587 | T688 | T689 | arg1Of | proliferation,[ |
| R588 | T688 | T692 | arg1Of | proliferation,"," |
| R589 | T690 | T689 | arg2Of | 13–16,[ |
| R590 | T691 | T689 | arg3Of | ],[ |
| R591 | T692 | T698 | arg1Of | ",",and |
| R592 | T694 | T692 | arg2Of | signals,"," |
| R593 | T694 | T693 | arg1Of | signals,anti-apoptotic |
| R594 | T694 | T695 | arg1Of | signals,[ |
| R595 | T696 | T695 | arg2Of | "17,18",[ |
| R596 | T697 | T695 | arg3Of | ],[ |
| R597 | T698 | T686 | arg2Of | and,including |
| R598 | T700 | T698 | arg2Of | development,and |
| R599 | T700 | T699 | arg1Of | development,thymocyte |
| R600 | T700 | T701 | arg1Of | development,[ |
| R601 | T702 | T701 | arg2Of | 19,[ |
| R602 | T703 | T701 | arg3Of | ],[ |
| R603 | T704 | T705 | arg1Of | Expression,of |
| R604 | T704 | T713 | arg1Of | Expression,can |
| R605 | T704 | T715 | arg1Of | Expression,modify |
| R606 | T708 | T709 | arg1Of | inactive,and |
| R607 | T710 | T709 | arg2Of | constitutively,and |
| R608 | T712 | T705 | arg2Of | PKDs,of |
| R609 | T712 | T706 | arg1Of | PKDs,mutant |
| R610 | T712 | T707 | arg1Of | PKDs,catalytically |
| R611 | T712 | T708 | arg1Of | PKDs,inactive |
| R612 | T712 | T710 | arg1Of | PKDs,constitutively |
| R613 | T712 | T711 | arg2Of | PKDs,activated |
| R614 | T715 | T713 | arg2Of | modify,can |
| R615 | T715 | T714 | arg1Of | modify,also |
| R616 | T717 | T716 | arg1Of | function,Golgi |
| R617 | T717 | T718 | arg1Of | function,"," |
| R618 | T718 | T721 | arg1Of | ",",and |
| R619 | T720 | T718 | arg2Of | adhesion,"," |
| R620 | T720 | T719 | arg1Of | adhesion,cell |
| R621 | T721 | T715 | arg2Of | and,modify |
| R622 | T723 | T721 | arg2Of | motility,and |
| R623 | T723 | T722 | arg1Of | motility,cell |
| R624 | T723 | T724 | arg1Of | motility,( |
| R625 | T725 | T724 | arg2Of | reviewed,( |
| R626 | T725 | T726 | arg1Of | reviewed,in |
| R627 | T725 | T727 | arg1Of | reviewed,[ |
| R628 | T728 | T727 | arg2Of | 20,[ |
| R629 | T729 | T727 | arg3Of | ],[ |
| R630 | T730 | T724 | arg3Of | ),( |
| R631 | T732 | T731 | arg2Of | particular,In |
| R632 | T734 | T735 | arg1Of | PKDs,have |
| R633 | T734 | T736 | arg1Of | PKDs,been |
| R634 | T734 | T738 | arg2Of | PKDs,linked |
| R635 | T734 | T749 | arg1Of | PKDs,regulating |
| R636 | T738 | T731 | arg1Of | linked,In |
| R637 | T738 | T733 | arg1Of | linked,"," |
| R638 | T738 | T735 | arg2Of | linked,have |
| R639 | T738 | T736 | arg2Of | linked,been |
| R640 | T738 | T737 | arg1Of | linked,widely |
| R641 | T738 | T739 | arg1Of | linked,to |
| R642 | T738 | T748 | arg1Of | linked,in |
| R643 | T739 | T747 | arg1Of | to,and |
| R644 | T741 | T739 | arg2Of | activation,to |
| R645 | T741 | T740 | arg1Of | activation,the |
| R646 | T741 | T742 | arg1Of | activation,of |
| R647 | T746 | T742 | arg2Of | factor,of |
| R648 | T746 | T743 | arg1Of | factor,the |
| R649 | T746 | T744 | arg1Of | factor,NFκB |
| R650 | T746 | T745 | arg1Of | factor,transcription |
| R651 | T748 | T747 | arg2Of | in,and |
| R652 | T749 | T748 | arg2Of | regulating,in |
| R653 | T749 | T752 | arg1Of | regulating,during |
| R654 | T751 | T749 | arg2Of | survival,regulating |
| R655 | T751 | T750 | arg1Of | survival,cell |
| R656 | T754 | T752 | arg2Of | stress,during |
| R657 | T754 | T753 | arg1Of | stress,oxidative |
| R658 | T754 | T755 | arg1Of | stress,[ |
| R659 | T756 | T755 | arg2Of | "17,21–23",[ |
| R660 | T757 | T755 | arg3Of | ],[ |
| R661 | T760 | T759 | arg1Of | proposed,recently |
| R662 | T762 | T758 | arg1Of | substrate,Another |
| R663 | T762 | T760 | arg1Of | substrate,proposed |
| R664 | T762 | T761 | arg1Of | substrate,PKD1 |
| R665 | T762 | T763 | arg1Of | substrate,is |
| R666 | T764 | T763 | arg2Of | HSP27,is |
| R667 | T764 | T765 | arg1Of | HSP27,[ |
| R668 | T764 | T768 | arg1Of | HSP27,"," |
| R669 | T766 | T765 | arg2Of | 24,[ |
| R670 | T767 | T765 | arg3Of | ],[ |
| R671 | T773 | T768 | arg2Of | protein,"," |
| R672 | T773 | T769 | arg1Of | protein,a |
| R673 | T773 | T770 | arg1Of | protein,small |
| R674 | T773 | T771 | arg1Of | protein,heat |
| R675 | T773 | T772 | arg1Of | protein,shock |
| R676 | T773 | T774 | arg2Of | protein,involved |
| R677 | T774 | T775 | arg1Of | involved,in |
| R678 | T776 | T775 | arg2Of | regulating,in |
| R679 | T776 | T782 | arg1Of | regulating,[ |
| R680 | T778 | T777 | arg1Of | migration,cell |
| R681 | T778 | T779 | arg1Of | migration,and |
| R682 | T779 | T776 | arg2Of | and,regulating |
| R683 | T781 | T779 | arg2Of | survival,and |
| R684 | T781 | T780 | arg1Of | survival,cell |
| R685 | T783 | T782 | arg2Of | 25,[ |
| R686 | T784 | T782 | arg3Of | ],[ |
| R687 | T787 | T785 | arg1Of | role,An |
| R688 | T787 | T786 | arg1Of | role,essential |
| R689 | T787 | T788 | arg1Of | role,for |
| R690 | T787 | T808 | arg1Of | role,has |
| R691 | T787 | T810 | arg1Of | role,been |
| R692 | T787 | T811 | arg2Of | role,demonstrated |
| R693 | T790 | T788 | arg2Of | enzymes,for |
| R694 | T790 | T789 | arg1Of | enzymes,PKD |
| R695 | T790 | T791 | arg1Of | enzymes,in |
| R696 | T792 | T791 | arg2Of | regulating,in |
| R697 | T796 | T792 | arg2Of | deacetylases,regulating |
| R698 | T796 | T793 | arg1Of | deacetylases,class |
| R699 | T796 | T794 | arg1Of | deacetylases,II |
| R700 | T796 | T795 | arg1Of | deacetylases,histone |
| R701 | T796 | T797 | arg1Of | deacetylases,( |
| R702 | T796 | T800 | arg1Of | deacetylases,"," |
| R703 | T798 | T797 | arg2Of | HDACs,( |
| R704 | T799 | T797 | arg3Of | ),( |
| R705 | T801 | T800 | arg2Of | enzymes,"," |
| R706 | T801 | T802 | arg1Of | enzymes,that |
| R707 | T801 | T803 | arg1Of | enzymes,repress |
| R708 | T806 | T803 | arg2Of | transcription,repress |
| R709 | T806 | T804 | arg1Of | transcription,MEF2-dependent |
| R710 | T806 | T805 | arg1Of | transcription,gene |
| R711 | T811 | T807 | arg1Of | demonstrated,"," |
| R712 | T811 | T808 | arg2Of | demonstrated,has |
| R713 | T811 | T809 | arg1Of | demonstrated,also |
| R714 | T811 | T810 | arg2Of | demonstrated,been |
| R715 | T811 | T812 | arg1Of | demonstrated,[ |
| R716 | T813 | T812 | arg2Of | "1,26–28",[ |
| R717 | T814 | T812 | arg3Of | ],[ |
| R718 | T816 | T815 | arg1Of | investigate,To |
| R719 | T819 | T816 | arg2Of | role,investigate |
| R720 | T819 | T817 | arg1Of | role,the |
| R721 | T819 | T818 | arg1Of | role,biological |
| R722 | T819 | T820 | arg1Of | role,of |
| R723 | T821 | T820 | arg2Of | PKDs,of |
| R724 | T822 | T816 | arg1Of | we,investigate |
| R725 | T822 | T823 | arg1Of | we,have |
| R726 | T822 | T824 | arg1Of | we,generated |
| R727 | T824 | T815 | modOf | generated,To |
| R728 | T824 | T823 | arg2Of | generated,have |
| R729 | T828 | T824 | arg2Of | lines,generated |
| R730 | T828 | T825 | arg1Of | lines,DT40 |
| R731 | T828 | T826 | arg1Of | lines,B |
| R732 | T828 | T827 | arg1Of | lines,cell |
| R733 | T828 | T829 | arg1Of | lines,that |
| R734 | T828 | T830 | arg1Of | lines,lack |
| R735 | T830 | T844 | arg1Of | lack,"," |
| R736 | T830 | T845 | modOf | lack,allowing |
| R737 | T830 | T866 | arg1Of | lack,"," |
| R738 | T830 | T869 | modOf | lack,addressing |
| R739 | T831 | T830 | arg2Of | expression,lack |
| R740 | T831 | T832 | arg1Of | expression,of |
| R741 | T833 | T834 | arg1Of | one,or |
| R742 | T833 | T835 | arg1Of | one,more |
| R743 | T836 | T832 | arg2Of | members,of |
| R744 | T836 | T833 | arg1Of | members,one |
| R745 | T836 | T837 | arg1Of | members,of |
| R746 | T840 | T837 | arg2Of | family,of |
| R747 | T840 | T838 | arg1Of | family,the |
| R748 | T840 | T839 | arg1Of | family,PKD |
| R749 | T840 | T841 | arg1Of | family,[ |
| R750 | T842 | T841 | arg2Of | 1,[ |
| R751 | T843 | T841 | arg3Of | ],[ |
| R752 | T846 | T845 | arg2Of | us,allowing |
| R753 | T846 | T848 | arg1Of | us,investigate |
| R754 | T848 | T845 | arg3Of | investigate,allowing |
| R755 | T848 | T847 | arg1Of | investigate,to |
| R756 | T850 | T848 | arg2Of | function,investigate |
| R757 | T850 | T849 | arg1Of | function,the |
| R758 | T850 | T851 | arg1Of | function,( |
| R759 | T850 | T854 | arg1Of | function,of |
| R760 | T850 | T857 | arg1Of | function,following |
| R761 | T852 | T851 | arg2Of | s,( |
| R762 | T853 | T851 | arg3Of | ),( |
| R763 | T856 | T854 | arg2Of | isoforms,of |
| R764 | T856 | T855 | arg1Of | isoforms,PKD |
| R765 | T863 | T862 | arg2Of | BCR,( |
| R766 | T864 | T862 | arg3Of | ),( |
| R767 | T865 | T857 | arg2Of | stimulation,following |
| R768 | T865 | T858 | arg1Of | stimulation,B |
| R769 | T865 | T859 | arg1Of | stimulation,cell |
| R770 | T865 | T860 | arg1Of | stimulation,antigen |
| R771 | T865 | T861 | arg1Of | stimulation,receptor |
| R772 | T865 | T862 | arg1Of | stimulation,( |
| R773 | T869 | T867 | arg1Of | addressing,as |
| R774 | T869 | T868 | arg1Of | addressing,well |
| R775 | T871 | T869 | arg2Of | issue,addressing |
| R776 | T871 | T870 | arg1Of | issue,the |
| R777 | T871 | T872 | arg1Of | issue,of |
| R778 | T874 | T872 | arg2Of | redundancy,of |
| R779 | T874 | T873 | arg1Of | redundancy,functional |
| R780 | T874 | T875 | arg1Of | redundancy,between |
| R781 | T880 | T875 | arg2Of | members,between |
| R782 | T880 | T876 | arg1Of | members,the |
| R783 | T880 | T877 | arg1Of | members,different |
| R784 | T880 | T878 | arg1Of | members,PKD |
| R785 | T880 | T879 | arg1Of | members,family |
| R786 | T882 | T881 | arg1Of | studies,Previous |
| R787 | T882 | T883 | arg1Of | studies,have |
| R788 | T882 | T884 | arg1Of | studies,shown |
| R789 | T884 | T883 | arg2Of | shown,have |
| R790 | T886 | T887 | arg1Of | PKDs,are |
| R791 | T886 | T888 | arg1Of | PKDs,indispensable |
| R792 | T887 | T884 | arg2Of | are,shown |
| R793 | T887 | T885 | arg1Of | are,that |
| R794 | T887 | T895 | arg1Of | are,[ |
| R795 | T888 | T887 | arg2Of | indispensable,are |
| R796 | T888 | T889 | arg1Of | indispensable,for |
| R797 | T891 | T889 | arg2Of | regulation,for |
| R798 | T891 | T890 | arg1Of | regulation,HDAC |
| R799 | T891 | T892 | arg1Of | regulation,in |
| R800 | T894 | T892 | arg2Of | cells,in |
| R801 | T894 | T893 | arg1Of | cells,B |
| R802 | T896 | T895 | arg2Of | 1,[ |
| R803 | T897 | T895 | arg3Of | ],[ |
| R804 | T899 | T900 | arg1Of | we,show |
| R805 | T900 | T898 | arg1Of | show,Herein |
| R806 | T902 | T903 | arg1Of | PKDs,are |
| R807 | T902 | T905 | arg1Of | PKDs,indispensable |
| R808 | T903 | T900 | arg2Of | are,show |
| R809 | T903 | T901 | arg1Of | are,that |
| R810 | T903 | T904 | arg1Of | are,also |
| R811 | T905 | T903 | arg2Of | indispensable,are |
| R812 | T905 | T906 | arg1Of | indispensable,for |
| R813 | T908 | T906 | arg2Of | phosphorylation,for |
| R814 | T908 | T907 | arg1Of | phosphorylation,HSP27 |
| R815 | T908 | T909 | arg1Of | phosphorylation,in |
| R816 | T911 | T909 | arg2Of | cells,in |
| R817 | T911 | T910 | arg1Of | cells,B |
| R818 | T917 | T914 | arg1Of | cells,PKD-null |
| R819 | T917 | T915 | arg1Of | cells,DT40 |
| R820 | T917 | T916 | arg1Of | cells,B |
| R821 | T917 | T918 | arg1Of | cells,are |
| R822 | T917 | T919 | arg1Of | cells,viable |
| R823 | T917 | T921 | arg1Of | cells,proliferate |
| R824 | T918 | T920 | arg1Of | are,and |
| R825 | T919 | T918 | arg2Of | viable,are |
| R826 | T920 | T912 | arg1Of | and,However |
| R827 | T920 | T913 | arg1Of | and,"," |
| R828 | T921 | T920 | arg2Of | proliferate,and |
| R829 | T921 | T922 | arg1Of | proliferate,normally |
| R830 | T925 | T926 | arg1Of | loss,of |
| R831 | T925 | T933 | arg1Of | loss,does |
| R832 | T925 | T936 | arg1Of | loss,affect |
| R833 | T925 | T944 | arg1Of | loss,do |
| R834 | T930 | T926 | arg2Of | pool,of |
| R835 | T930 | T927 | arg1Of | pool,the |
| R836 | T930 | T928 | arg1Of | pool,entire |
| R837 | T930 | T929 | arg1Of | pool,cellular |
| R838 | T930 | T931 | arg1Of | pool,of |
| R839 | T932 | T931 | arg2Of | PKD,of |
| R840 | T936 | T933 | arg2Of | affect,does |
| R841 | T936 | T934 | arg1Of | affect,not |
| R842 | T936 | T935 | arg1Of | affect,critically |
| R843 | T936 | T943 | arg1Of | affect,nor |
| R844 | T939 | T936 | arg2Of | responses,affect |
| R845 | T939 | T937 | arg1Of | responses,oxidative |
| R846 | T939 | T938 | arg1Of | responses,stress |
| R847 | T939 | T940 | arg1Of | responses,in |
| R848 | T942 | T940 | arg2Of | cells,in |
| R849 | T942 | T941 | arg1Of | cells,B |
| R850 | T943 | T923 | arg1Of | nor,Moreover |
| R851 | T943 | T924 | arg1Of | nor,"," |
| R852 | T944 | T943 | arg2Of | do,nor |
| R853 | T945 | T944 | arg2Of | PKD,do |
| R854 | T946 | T947 | arg1Of | kinases,play |
| R855 | T946 | T952 | arg1Of | kinases,regulating |
| R856 | T947 | T944 | arg3Of | play,do |
| R857 | T947 | T951 | arg1Of | play,in |
| R858 | T950 | T947 | arg2Of | role,play |
| R859 | T950 | T948 | arg1Of | role,an |
| R860 | T950 | T949 | arg1Of | role,essential |
| R861 | T952 | T951 | arg2Of | regulating,in |
| R862 | T955 | T952 | arg2Of | activity,regulating |
| R863 | T955 | T953 | arg1Of | activity,NFκB |
| R864 | T955 | T954 | arg1Of | activity,transcriptional |
| R865 | T959 | T958 | arg1Of | findings,these |
| R866 | T959 | T960 | arg1Of | findings,reveal |
| R867 | T960 | T956 | arg1Of | reveal,Together |
| R868 | T960 | T957 | arg1Of | reveal,"," |
| R869 | T964 | T962 | arg2Of | lymphocytes,in |
| R870 | T964 | T963 | arg1Of | lymphocytes,B |
| R871 | T967 | T966 | arg1Of | kinases,PKD |
| R872 | T967 | T968 | arg1Of | kinases,are |
| R873 | T968 | T960 | arg2Of | are,reveal |
| R874 | T968 | T961 | arg1Of | are,that |
| R875 | T968 | T962 | arg1Of | are,in |
| R876 | T968 | T965 | arg1Of | are,"," |
| R877 | T968 | T969 | arg1Of | are,not |
| R878 | T971 | T968 | arg2Of | regulators,are |
| R879 | T971 | T970 | arg1Of | regulators,critical |
| R880 | T971 | T972 | arg1Of | regulators,of |
| R881 | T973 | T972 | arg2Of | many,of |
| R882 | T973 | T974 | arg1Of | many,of |
| R883 | T977 | T974 | arg2Of | processes,of |
| R884 | T977 | T975 | arg1Of | processes,the |
| R885 | T977 | T976 | arg1Of | processes,cellular |
| R886 | T977 | T979 | arg2Of | processes,ascribed |
| R887 | T979 | T978 | arg1Of | ascribed,previously |
| R888 | T979 | T980 | arg1Of | ascribed,to |
| R889 | T979 | T982 | arg1Of | ascribed,in |
| R890 | T981 | T980 | arg2Of | them,to |
| R891 | T985 | T982 | arg2Of | systems,in |
| R892 | T985 | T983 | arg1Of | systems,other |
| R893 | T985 | T984 | arg1Of | systems,cellular |
| R1437 | T1675 | T1674 | arg1Of | culture,"Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell" |
| R1438 | T1675 | T1676 | arg1Of | culture,"," |
| R1439 | T1676 | T1679 | arg1Of | ",",and |
| R1440 | T1678 | T1676 | arg2Of | transfections,"," |
| R1441 | T1678 | T1677 | arg1Of | transfections,transient |
| R1442 | T1681 | T1679 | arg2Of | stimulation,and |
| R1443 | T1681 | T1680 | arg1Of | stimulation,cell |
| R1444 | T1683 | T1684 | arg1Of | generation,"," |
| R1445 | T1684 | T1686 | arg1Of | ",",and |
| R1446 | T1685 | T1684 | arg2Of | culture,"," |
| R1447 | T1686 | T1682 | arg1Of | and,The |
| R1448 | T1686 | T1688 | arg1Of | and,of |
| R1449 | T1686 | T1699 | arg1Of | and,have |
| R1450 | T1686 | T1700 | arg1Of | and,been |
| R1451 | T1686 | T1701 | arg2Of | and,described |
| R1452 | T1687 | T1686 | arg2Of | activation,and |
| R1453 | T1689 | T1690 | arg1Of | PKD1−/−,"," |
| R1454 | T1690 | T1692 | arg1Of | ",",and |
| R1455 | T1691 | T1690 | arg2Of | PKD3−/−,"," |
| R1456 | T1693 | T1692 | arg2Of | PKD1/3−/−,and |
| R1457 | T1698 | T1688 | arg2Of | lines,of |
| R1458 | T1698 | T1689 | arg1Of | lines,PKD1−/− |
| R1459 | T1698 | T1691 | arg1Of | lines,PKD3−/− |
| R1460 | T1698 | T1693 | arg1Of | lines,PKD1/3−/− |
| R1461 | T1698 | T1694 | arg1Of | lines,knockout |
| R1462 | T1698 | T1695 | arg1Of | lines,DT40 |
| R1463 | T1698 | T1696 | arg1Of | lines,B |
| R1464 | T1698 | T1697 | arg1Of | lines,cell |
| R1465 | T1701 | T1699 | arg2Of | described,have |
| R1466 | T1701 | T1700 | arg2Of | described,been |
| R1467 | T1701 | T1702 | arg1Of | described,previously |
| R1468 | T1701 | T1703 | arg1Of | described,[ |
| R1469 | T1704 | T1703 | arg2Of | 1,[ |
| R1470 | T1705 | T1703 | arg3Of | ],[ |
| R1471 | T1706 | T1707 | arg1Of | Cells,were |
| R1472 | T1706 | T1708 | arg2Of | Cells,lysed |
| R1473 | T1708 | T1707 | arg2Of | lysed,were |
| R1474 | T1708 | T1709 | arg1Of | lysed,and |
| R1475 | T1711 | T1710 | arg1Of | extracts,protein |
| R1476 | T1711 | T1712 | arg1Of | extracts,were |
| R1477 | T1711 | T1713 | arg2Of | extracts,analysed |
| R1478 | T1713 | T1709 | arg2Of | analysed,and |
| R1479 | T1713 | T1712 | arg2Of | analysed,were |
| R1480 | T1713 | T1714 | arg1Of | analysed,in |
| R1481 | T1713 | T1718 | arg1Of | analysed,as |
| R1482 | T1717 | T1714 | arg2Of | experiments,in |
| R1483 | T1717 | T1715 | arg1Of | experiments,Western |
| R1484 | T1717 | T1716 | arg1Of | experiments,blotting |
| R1485 | T1720 | T1719 | arg1Of | described,previously |
| R1486 | T1722 | T1718 | arg2Of | 1,as |
| R1487 | T1722 | T1720 | arg1Of | 1,described |
| R1488 | T1722 | T1721 | arg2Of | 1,[ |
| R1489 | T1723 | T1721 | arg3Of | ],[ |
| R1490 | T1727 | T1724 | arg1Of | assays,Chloramphenicol |
| R1491 | T1727 | T1725 | arg1Of | assays,acetyl |
| R1492 | T1727 | T1726 | arg1Of | assays,transferase |
| R1493 | T1727 | T1728 | arg1Of | assays,have |
| R1494 | T1727 | T1729 | arg1Of | assays,been |
| R1495 | T1727 | T1730 | arg2Of | assays,described |
| R1496 | T1730 | T1728 | arg2Of | described,have |
| R1497 | T1730 | T1729 | arg2Of | described,been |
| R1498 | T1730 | T1731 | arg1Of | described,previously |
| R1499 | T1730 | T1732 | arg1Of | described,[ |
| R1500 | T1733 | T1732 | arg2Of | 29,[ |
| R1501 | T1734 | T1732 | arg3Of | ],[ |
| R1596 | T1869 | T1868 | arg1Of | staining,"Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell culture, transient transfections and cell stimulation The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. Chloramphenicol acetyl transferase assays have been described previously [29]. 2.2 sIgM" |
| R7 | T51 | T49 | arg1Of | D,protein |
| R8 | T51 | T50 | arg1Of | D,kinase |
| R12 | T55 | T48 | arg2Of | enzymes,of |
| R13 | T55 | T51 | arg1Of | enzymes,D |
| R14 | T56 | T45 | arg1Of | we,investigate |
| R16 | T57 | T44 | modOf | generated,To |
| R374 | T29 | T26 | arg1Of | enzymes,Protein |
| R375 | T29 | T27 | arg1Of | enzymes,kinase |
| R376 | T29 | T28 | arg1Of | enzymes,D |
| R377 | T29 | T30 | arg1Of | enzymes,are |
| R378 | T29 | T31 | arg1Of | enzymes,dispensable |
| R379 | T31 | T30 | arg2Of | dispensable,are |
| R380 | T31 | T32 | arg1Of | dispensable,for |
| R381 | T33 | T34 | arg1Of | proliferation,"," |
| R382 | T34 | T36 | arg1Of | ",",and |
| R383 | T35 | T34 | arg2Of | survival,"," |
| R384 | T36 | T32 | arg2Of | and,for |
| R385 | T36 | T41 | arg1Of | and,in |
| R386 | T40 | T36 | arg2Of | activity,and |
| R387 | T40 | T37 | arg1Of | activity,antigen |
| R388 | T40 | T38 | arg1Of | activity,receptor-regulated |
| R389 | T40 | T39 | arg1Of | activity,NFκB |
| R390 | T43 | T41 | arg2Of | B-cells,in |
| R391 | T43 | T42 | arg1Of | B-cells,vertebrate |
| R392 | T45 | T44 | arg1Of | investigate,To |
| R393 | T47 | T45 | arg2Of | importance,investigate |
| R394 | T47 | T46 | arg1Of | importance,the |
| R395 | T47 | T48 | arg1Of | importance,of |
bionlp-st-ge-2016-spacy-parsed
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T212 | 0-7 | NN | denotes | Protein |
| T213 | 8-14 | NN | denotes | kinase |
| T214 | 15-16 | NNP | denotes | D |
| T215 | 17-24 | NNS | denotes | enzymes |
| T216 | 25-28 | VBP | denotes | are |
| T217 | 29-40 | JJ | denotes | dispensable |
| T218 | 41-44 | IN | denotes | for |
| T219 | 45-58 | NN | denotes | proliferation |
| T220 | 58-59 | , | denotes | , |
| T221 | 60-68 | NN | denotes | survival |
| T222 | 69-72 | CC | denotes | and |
| T223 | 73-80 | NN | denotes | antigen |
| T224 | 81-99 | JJ | denotes | receptor-regulated |
| T225 | 100-104 | NN | denotes | NFκB |
| T226 | 105-113 | NN | denotes | activity |
| T227 | 114-116 | IN | denotes | in |
| T228 | 117-127 | JJ | denotes | vertebrate |
| T229 | 128-135 | NNPS | denotes | B-cells |
| T230 | 147-149 | TO | denotes | To |
| T231 | 150-161 | VB | denotes | investigate |
| T232 | 162-165 | DT | denotes | the |
| T233 | 166-176 | NN | denotes | importance |
| T234 | 177-179 | IN | denotes | of |
| T235 | 180-187 | NN | denotes | protein |
| T236 | 188-194 | NN | denotes | kinase |
| T237 | 195-196 | NNP | denotes | D |
| T238 | 197-198 | -LRB- | denotes | ( |
| T239 | 198-201 | NNP | denotes | PKD |
| T240 | 201-202 | -RRB- | denotes | ) |
| T241 | 203-210 | VBZ | denotes | enzymes |
| T242 | 211-213 | PRP | denotes | we |
| T243 | 214-223 | VBD | denotes | generated |
| T244 | 224-225 | DT | denotes | a |
| T245 | 226-234 | NNP | denotes | PKD-null |
| T246 | 235-239 | NNP | denotes | DT40 |
| T247 | 240-252 | NNP | denotes | B-lymphocyte |
| T248 | 253-257 | NN | denotes | cell |
| T249 | 258-262 | NN | denotes | line |
| T250 | 262-263 | . | denotes | . |
| T251 | 264-274 | RB | denotes | Previously |
| T252 | 275-277 | PRP | denotes | we |
| T253 | 278-282 | VBP | denotes | have |
| T254 | 283-288 | VBN | denotes | shown |
| T255 | 289-293 | IN | denotes | that |
| T256 | 294-298 | NNS | denotes | PKDs |
| T257 | 299-303 | VBP | denotes | have |
| T258 | 304-306 | DT | denotes | an |
| T259 | 307-316 | JJ | denotes | essential |
| T260 | 317-321 | NN | denotes | role |
| T261 | 322-324 | IN | denotes | in |
| T262 | 325-335 | VBG | denotes | regulating |
| T263 | 336-341 | NN | denotes | class |
| T264 | 342-344 | NNP | denotes | II |
| T265 | 345-352 | NN | denotes | histone |
| T266 | 353-365 | NNS | denotes | deacetylases |
| T267 | 366-368 | IN | denotes | in |
| T268 | 369-373 | NNP | denotes | DT40 |
| T269 | 374-381 | NNPS | denotes | B-cells |
| T270 | 382-383 | NNP | denotes | [ |
| T271 | 383-391 | NNP | denotes | Matthews |
| T272 | 391-392 | , | denotes | , |
| T273 | 393-397 | NNP | denotes | S.A. |
| T274 | 397-398 | , | denotes | , |
| T275 | 399-402 | NNP | denotes | Liu |
| T276 | 402-403 | , | denotes | , |
| T277 | 404-406 | NNP | denotes | P. |
| T278 | 406-407 | , | denotes | , |
| T279 | 408-416 | NNP | denotes | Spitaler |
| T280 | 416-417 | , | denotes | , |
| T281 | 418-420 | NNP | denotes | M. |
| T282 | 420-421 | , | denotes | , |
| T283 | 422-427 | NNP | denotes | Olson |
| T284 | 427-428 | , | denotes | , |
| T285 | 429-433 | NNP | denotes | E.N. |
| T286 | 433-434 | , | denotes | , |
| T287 | 435-443 | NNP | denotes | McKinsey |
| T288 | 443-444 | , | denotes | , |
| T289 | 445-449 | NNP | denotes | T.A. |
| T290 | 449-450 | , | denotes | , |
| T291 | 451-459 | NNP | denotes | Cantrell |
| T292 | 459-460 | , | denotes | , |
| T293 | 461-465 | NNP | denotes | D.A. |
| T294 | 466-469 | CC | denotes | and |
| T295 | 470-481 | NNP | denotes | Scharenberg |
| T296 | 481-482 | , | denotes | , |
| T297 | 483-487 | NNP | denotes | A.M. |
| T298 | 488-489 | -LRB- | denotes | ( |
| T299 | 489-493 | CD | denotes | 2006 |
| T300 | 493-494 | -RRB- | denotes | ) |
| T301 | 495-504 | JJ | denotes | Essential |
| T302 | 505-509 | NN | denotes | role |
| T303 | 510-513 | IN | denotes | for |
| T304 | 514-521 | NN | denotes | protein |
| T305 | 522-528 | NN | denotes | kinase |
| T306 | 529-530 | NNP | denotes | D |
| T307 | 531-537 | NN | denotes | family |
| T308 | 538-545 | NNS | denotes | kinases |
| T309 | 546-548 | IN | denotes | in |
| T310 | 549-552 | DT | denotes | the |
| T311 | 553-563 | NN | denotes | regulation |
| T312 | 564-566 | IN | denotes | of |
| T313 | 567-572 | NN | denotes | class |
| T314 | 573-575 | NNP | denotes | II |
| T315 | 576-583 | NN | denotes | histone |
| T316 | 584-596 | NNS | denotes | deacetylases |
| T317 | 597-599 | IN | denotes | in |
| T318 | 600-601 | NNP | denotes | B |
| T319 | 602-613 | NNS | denotes | lymphocytes |
| T320 | 613-614 | . | denotes | . |
| T321 | 615-618 | NNP | denotes | Mol |
| T322 | 618-619 | . | denotes | . |
| T323 | 620-624 | NNP | denotes | Cell |
| T324 | 625-629 | NNP | denotes | Biol |
| T325 | 629-630 | . | denotes | . |
| T326 | 631-633 | CD | denotes | 26 |
| T327 | 633-634 | , | denotes | , |
| T328 | 635-639 | CD | denotes | 1569 |
| T329 | 640-644 | CD | denotes | 1577 |
| T330 | 644-645 | NNP | denotes | ] |
| T331 | 645-646 | . | denotes | . |
| T332 | 647-649 | PRP | denotes | We |
| T333 | 650-653 | RB | denotes | now |
| T334 | 654-658 | VBP | denotes | show |
| T335 | 659-663 | IN | denotes | that |
| T336 | 664-668 | NNS | denotes | PKDs |
| T337 | 669-672 | VBP | denotes | are |
| T338 | 673-677 | RB | denotes | also |
| T339 | 678-686 | VBN | denotes | required |
| T340 | 687-689 | TO | denotes | to |
| T341 | 690-698 | VB | denotes | regulate |
| T342 | 699-704 | NNP | denotes | HSP27 |
| T343 | 705-720 | NN | denotes | phosphorylation |
| T344 | 721-723 | IN | denotes | in |
| T345 | 724-728 | NNP | denotes | DT40 |
| T346 | 729-736 | NNPS | denotes | B-cells |
| T347 | 736-737 | . | denotes | . |
| T348 | 738-745 | RB | denotes | However |
| T349 | 745-746 | , | denotes | , |
| T350 | 747-749 | IN | denotes | in |
| T351 | 750-758 | NN | denotes | contrast |
| T352 | 759-761 | TO | denotes | to |
| T353 | 762-770 | JJ | denotes | previous |
| T354 | 771-783 | NNS | denotes | observations |
| T355 | 784-786 | IN | denotes | in |
| T356 | 787-792 | JJ | denotes | other |
| T357 | 793-797 | NN | denotes | cell |
| T358 | 798-803 | NNS | denotes | types |
| T359 | 803-804 | , | denotes | , |
| T360 | 805-808 | NNP | denotes | PKD |
| T361 | 809-816 | VBZ | denotes | enzymes |
| T362 | 817-819 | VB | denotes | do |
| T363 | 820-823 | RB | denotes | not |
| T364 | 824-832 | VB | denotes | regulate |
| T365 | 833-838 | JJ | denotes | basic |
| T366 | 839-847 | JJ | denotes | cellular |
| T367 | 848-857 | NNS | denotes | processes |
| T368 | 858-862 | JJ | denotes | such |
| T369 | 863-865 | IN | denotes | as |
| T370 | 866-879 | NN | denotes | proliferation |
| T371 | 880-882 | CC | denotes | or |
| T372 | 883-891 | NN | denotes | survival |
| T373 | 892-901 | NNS | denotes | responses |
| T374 | 901-902 | , | denotes | , |
| T375 | 903-906 | CC | denotes | nor |
| T376 | 907-911 | NNP | denotes | NFκB |
| T377 | 912-927 | JJ | denotes | transcriptional |
| T378 | 928-936 | NN | denotes | activity |
| T379 | 937-947 | JJ | denotes | downstream |
| T380 | 948-950 | IN | denotes | of |
| T381 | 951-954 | DT | denotes | the |
| T382 | 955-956 | NNP | denotes | B |
| T383 | 957-961 | NN | denotes | cell |
| T384 | 962-969 | NN | denotes | antigen |
| T385 | 970-978 | NN | denotes | receptor |
| T386 | 978-979 | . | denotes | . |
| T387 | 980-984 | RB | denotes | Thus |
| T388 | 984-985 | , | denotes | , |
| T389 | 986-990 | NNS | denotes | PKDs |
| T390 | 991-995 | VBP | denotes | have |
| T391 | 996-997 | DT | denotes | a |
| T392 | 998-1007 | JJ | denotes | selective |
| T393 | 1008-1012 | NN | denotes | role |
| T394 | 1013-1015 | IN | denotes | in |
| T395 | 1016-1020 | NNP | denotes | DT40 |
| T396 | 1021-1027 | NNP | denotes | B-cell |
| T397 | 1028-1035 | NN | denotes | biology |
| T398 | 1035-1036 | . | denotes | . |
| T1008 | 635-1057 | JJ | denotes | 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The |
| T1009 | 1058-1065 | NN | denotes | protein |
| T1010 | 1066-1072 | NN | denotes | kinase |
| T1011 | 1073-1074 | NNP | denotes | D |
| T1012 | 1075-1076 | -LRB- | denotes | ( |
| T1013 | 1076-1079 | NNP | denotes | PKD |
| T1014 | 1079-1080 | -RRB- | denotes | ) |
| T1015 | 1081-1097 | JJ | denotes | serine/threonine |
| T1016 | 1098-1104 | NN | denotes | kinase |
| T1017 | 1105-1111 | NN | denotes | family |
| T1018 | 1112-1115 | VBZ | denotes | has |
| T1019 | 1116-1121 | CD | denotes | three |
| T1020 | 1122-1129 | NNS | denotes | members |
| T1021 | 1129-1130 | : | denotes | : |
| T1022 | 1131-1135 | NNP | denotes | PKD1 |
| T1023 | 1135-1136 | , | denotes | , |
| T1024 | 1137-1141 | NNP | denotes | PKD2 |
| T1025 | 1142-1145 | CC | denotes | and |
| T1026 | 1146-1150 | NNP | denotes | PKD3 |
| T1027 | 1150-1151 | . | denotes | . |
| T1028 | 1152-1156 | JJS | denotes | Most |
| T1029 | 1157-1161 | NN | denotes | cell |
| T1030 | 1162-1167 | NNS | denotes | types |
| T1031 | 1168-1175 | VBP | denotes | express |
| T1032 | 1176-1178 | IN | denotes | at |
| T1033 | 1179-1184 | JJS | denotes | least |
| T1034 | 1185-1188 | CD | denotes | two |
| T1035 | 1189-1192 | NNP | denotes | PKD |
| T1036 | 1193-1201 | NNS | denotes | isoforms |
| T1037 | 1202-1205 | CC | denotes | but |
| T1038 | 1206-1209 | NNP | denotes | PKD |
| T1039 | 1210-1217 | NNS | denotes | enzymes |
| T1040 | 1218-1221 | VBP | denotes | are |
| T1041 | 1222-1232 | RB | denotes | especially |
| T1042 | 1233-1239 | RB | denotes | highly |
| T1043 | 1240-1249 | VBN | denotes | expressed |
| T1044 | 1250-1252 | IN | denotes | in |
| T1045 | 1253-1267 | JJ | denotes | haematopoietic |
| T1046 | 1268-1273 | NNS | denotes | cells |
| T1047 | 1273-1274 | , | denotes | , |
| T1048 | 1275-1280 | WRB | denotes | where |
| T1049 | 1281-1285 | PRP | denotes | they |
| T1050 | 1286-1289 | VBP | denotes | are |
| T1051 | 1290-1299 | VBN | denotes | activated |
| T1052 | 1300-1302 | IN | denotes | in |
| T1053 | 1303-1311 | NN | denotes | response |
| T1054 | 1312-1314 | TO | denotes | to |
| T1055 | 1315-1322 | NN | denotes | antigen |
| T1056 | 1323-1332 | NNS | denotes | receptors |
| T1057 | 1333-1344 | NN | denotes | stimulation |
| T1058 | 1345-1346 | NNP | denotes | [ |
| T1059 | 1346-1349 | CD | denotes | 2,3 |
| T1060 | 1349-1350 | NNP | denotes | ] |
| T1061 | 1350-1351 | . | denotes | . |
| T1062 | 1352-1353 | DT | denotes | A |
| T1063 | 1354-1363 | JJ | denotes | conserved |
| T1064 | 1364-1374 | JJ | denotes | signalling |
| T1065 | 1375-1382 | NN | denotes | pathway |
| T1066 | 1383-1390 | VBG | denotes | linking |
| T1067 | 1391-1398 | NN | denotes | antigen |
| T1068 | 1399-1408 | NNS | denotes | receptors |
| T1069 | 1409-1411 | TO | denotes | to |
| T1070 | 1412-1416 | NNS | denotes | PKDs |
| T1071 | 1417-1425 | VBZ | denotes | involves |
| T1072 | 1426-1429 | DT | denotes | the |
| T1073 | 1430-1440 | NN | denotes | activation |
| T1074 | 1441-1443 | IN | denotes | of |
| T1075 | 1444-1448 | NNP | denotes | PLCγ |
| T1076 | 1449-1452 | CC | denotes | and |
| T1077 | 1453-1456 | DT | denotes | the |
| T1078 | 1457-1467 | JJ | denotes | subsequent |
| T1079 | 1468-1478 | NN | denotes | production |
| T1080 | 1479-1481 | IN | denotes | of |
| T1081 | 1482-1496 | NN | denotes | diacylglycerol |
| T1082 | 1497-1498 | -LRB- | denotes | ( |
| T1083 | 1498-1501 | NNP | denotes | DAG |
| T1084 | 1501-1502 | -RRB- | denotes | ) |
| T1085 | 1503-1508 | WDT | denotes | which |
| T1086 | 1509-1519 | VBZ | denotes | stimulates |
| T1087 | 1520-1529 | JJ | denotes | classical |
| T1088 | 1530-1536 | CC | denotes | and/or |
| T1089 | 1537-1542 | JJ | denotes | novel |
| T1090 | 1543-1550 | NN | denotes | protein |
| T1091 | 1551-1557 | NN | denotes | kinase |
| T1092 | 1558-1560 | NNS | denotes | Cs |
| T1093 | 1561-1562 | -LRB- | denotes | ( |
| T1094 | 1562-1565 | NNP | denotes | PKC |
| T1095 | 1565-1566 | -RRB- | denotes | ) |
| T1096 | 1567-1571 | WDT | denotes | that |
| T1097 | 1572-1585 | VBP | denotes | phosphorylate |
| T1098 | 1586-1589 | CD | denotes | two |
| T1099 | 1590-1593 | JJ | denotes | key |
| T1100 | 1594-1604 | JJ | denotes | regulatory |
| T1101 | 1605-1611 | NN | denotes | serine |
| T1102 | 1612-1620 | NNS | denotes | residues |
| T1103 | 1621-1623 | IN | denotes | in |
| T1104 | 1624-1627 | DT | denotes | the |
| T1105 | 1628-1638 | NN | denotes | activation |
| T1106 | 1639-1643 | NN | denotes | loop |
| T1107 | 1644-1646 | IN | denotes | of |
| T1108 | 1647-1650 | NNP | denotes | PKD |
| T1109 | 1651-1658 | VBZ | denotes | kinases |
| T1110 | 1659-1660 | NNP | denotes | [ |
| T1111 | 1660-1661 | CD | denotes | 3 |
| T1112 | 1662-1663 | CD | denotes | 6 |
| T1113 | 1663-1664 | NNP | denotes | ] |
| T1114 | 1664-1665 | . | denotes | . |
| T1115 | 1666-1669 | DT | denotes | The |
| T1116 | 1670-1680 | JJ | denotes | N-terminal |
| T1117 | 1681-1691 | JJ | denotes | regulatory |
| T1118 | 1692-1698 | NN | denotes | region |
| T1119 | 1699-1701 | IN | denotes | of |
| T1120 | 1702-1705 | NNP | denotes | PKD |
| T1121 | 1706-1713 | VBZ | denotes | enzymes |
| T1122 | 1714-1722 | VBZ | denotes | contains |
| T1123 | 1723-1724 | DT | denotes | a |
| T1124 | 1725-1728 | NNP | denotes | DAG |
| T1125 | 1729-1736 | JJ | denotes | binding |
| T1126 | 1737-1743 | NN | denotes | domain |
| T1127 | 1744-1747 | CC | denotes | and |
| T1128 | 1748-1754 | JJ | denotes | direct |
| T1129 | 1755-1762 | JJ | denotes | binding |
| T1130 | 1763-1765 | IN | denotes | of |
| T1131 | 1766-1769 | NNP | denotes | DAG |
| T1132 | 1770-1774 | RB | denotes | also |
| T1133 | 1775-1786 | VBZ | denotes | contributes |
| T1134 | 1787-1789 | TO | denotes | to |
| T1135 | 1790-1794 | CD | denotes | PKD1 |
| T1136 | 1795-1805 | NN | denotes | activation |
| T1137 | 1806-1807 | NN | denotes | [ |
| T1138 | 1807-1808 | CD | denotes | 7 |
| T1139 | 1808-1809 | NNP | denotes | ] |
| T1140 | 1810-1812 | RB | denotes | as |
| T1141 | 1813-1817 | RB | denotes | well |
| T1142 | 1818-1820 | IN | denotes | as |
| T1143 | 1821-1831 | VBG | denotes | regulating |
| T1144 | 1832-1835 | DT | denotes | the |
| T1145 | 1836-1843 | JJ | denotes | spatial |
| T1146 | 1844-1852 | NN | denotes | location |
| T1147 | 1853-1855 | IN | denotes | of |
| T1148 | 1856-1859 | NNP | denotes | PKD |
| T1149 | 1860-1867 | VBZ | denotes | enzymes |
| T1150 | 1868-1874 | IN | denotes | within |
| T1151 | 1875-1880 | NNS | denotes | cells |
| T1152 | 1881-1882 | VBP | denotes | [ |
| T1153 | 1882-1883 | CD | denotes | 8 |
| T1154 | 1884-1886 | CD | denotes | 12 |
| T1155 | 1886-1887 | NN | denotes | ] |
| T1156 | 1887-1888 | . | denotes | . |
| T1157 | 1889-1892 | NNP | denotes | PKD |
| T1158 | 1893-1900 | NNS | denotes | enzymes |
| T1159 | 1901-1905 | VBP | denotes | have |
| T1160 | 1906-1910 | VBN | denotes | been |
| T1161 | 1911-1919 | VBN | denotes | proposed |
| T1162 | 1920-1922 | TO | denotes | to |
| T1163 | 1923-1931 | VB | denotes | regulate |
| T1164 | 1932-1940 | JJ | denotes | numerous |
| T1165 | 1941-1949 | JJ | denotes | cellular |
| T1166 | 1950-1959 | NNS | denotes | functions |
| T1167 | 1959-1960 | , | denotes | , |
| T1168 | 1961-1970 | VBG | denotes | including |
| T1169 | 1971-1975 | NN | denotes | cell |
| T1170 | 1976-1989 | NN | denotes | proliferation |
| T1171 | 1990-1991 | NNP | denotes | [ |
| T1172 | 1991-1993 | CD | denotes | 13 |
| T1173 | 1994-1996 | CD | denotes | 16 |
| T1174 | 1996-1997 | NNP | denotes | ] |
| T1175 | 1997-1998 | , | denotes | , |
| T1176 | 1999-2013 | JJ | denotes | anti-apoptotic |
| T1177 | 2014-2021 | NNS | denotes | signals |
| T1178 | 2022-2023 | NNP | denotes | [ |
| T1179 | 2023-2028 | CD | denotes | 17,18 |
| T1180 | 2028-2029 | NNP | denotes | ] |
| T1181 | 2030-2033 | CC | denotes | and |
| T1182 | 2034-2043 | JJ | denotes | thymocyte |
| T1183 | 2044-2055 | NN | denotes | development |
| T1184 | 2056-2057 | NNP | denotes | [ |
| T1185 | 2057-2059 | CD | denotes | 19 |
| T1186 | 2059-2060 | NNP | denotes | ] |
| T1187 | 2060-2061 | . | denotes | . |
| T1188 | 2062-2072 | NN | denotes | Expression |
| T1189 | 2073-2075 | IN | denotes | of |
| T1190 | 2076-2082 | JJ | denotes | mutant |
| T1191 | 2083-2096 | RB | denotes | catalytically |
| T1192 | 2097-2105 | JJ | denotes | inactive |
| T1193 | 2106-2109 | CC | denotes | and |
| T1194 | 2110-2124 | RB | denotes | constitutively |
| T1195 | 2125-2134 | VBN | denotes | activated |
| T1196 | 2135-2139 | NNS | denotes | PKDs |
| T1197 | 2140-2143 | MD | denotes | can |
| T1198 | 2144-2148 | RB | denotes | also |
| T1199 | 2149-2155 | VB | denotes | modify |
| T1200 | 2156-2161 | NNP | denotes | Golgi |
| T1201 | 2162-2170 | NN | denotes | function |
| T1202 | 2170-2171 | , | denotes | , |
| T1203 | 2172-2176 | NN | denotes | cell |
| T1204 | 2177-2185 | NN | denotes | adhesion |
| T1205 | 2186-2189 | CC | denotes | and |
| T1206 | 2190-2194 | NN | denotes | cell |
| T1207 | 2195-2203 | NN | denotes | motility |
| T1208 | 2204-2205 | -LRB- | denotes | ( |
| T1209 | 2205-2213 | VBN | denotes | reviewed |
| T1210 | 2214-2216 | IN | denotes | in |
| T1211 | 2217-2218 | NNP | denotes | [ |
| T1212 | 2218-2220 | CD | denotes | 20 |
| T1213 | 2220-2221 | NNP | denotes | ] |
| T1214 | 2221-2222 | -RRB- | denotes | ) |
| T1215 | 2222-2223 | . | denotes | . |
| T1216 | 2224-2226 | IN | denotes | In |
| T1217 | 2227-2237 | JJ | denotes | particular |
| T1218 | 2237-2238 | , | denotes | , |
| T1219 | 2239-2243 | NNS | denotes | PKDs |
| T1220 | 2244-2248 | VBP | denotes | have |
| T1221 | 2249-2253 | VBN | denotes | been |
| T1222 | 2254-2260 | RB | denotes | widely |
| T1223 | 2261-2267 | VBN | denotes | linked |
| T1224 | 2268-2270 | TO | denotes | to |
| T1225 | 2271-2274 | DT | denotes | the |
| T1226 | 2275-2285 | NN | denotes | activation |
| T1227 | 2286-2288 | IN | denotes | of |
| T1228 | 2289-2292 | DT | denotes | the |
| T1229 | 2293-2297 | NNP | denotes | NFκB |
| T1230 | 2298-2311 | NN | denotes | transcription |
| T1231 | 2312-2318 | NN | denotes | factor |
| T1232 | 2319-2322 | CC | denotes | and |
| T1233 | 2323-2325 | IN | denotes | in |
| T1234 | 2326-2336 | VBG | denotes | regulating |
| T1235 | 2337-2341 | NN | denotes | cell |
| T1236 | 2342-2350 | NN | denotes | survival |
| T1237 | 2351-2357 | IN | denotes | during |
| T1238 | 2358-2367 | JJ | denotes | oxidative |
| T1239 | 2368-2374 | NN | denotes | stress |
| T1240 | 2375-2376 | NNP | denotes | [ |
| T1241 | 2376-2381 | CD | denotes | 17,21 |
| T1242 | 2382-2384 | CD | denotes | 23 |
| T1243 | 2384-2385 | NNP | denotes | ] |
| T1244 | 2385-2386 | . | denotes | . |
| T1245 | 2387-2394 | DT | denotes | Another |
| T1246 | 2395-2403 | RB | denotes | recently |
| T1247 | 2404-2412 | VBN | denotes | proposed |
| T1248 | 2413-2417 | NNP | denotes | PKD1 |
| T1249 | 2418-2427 | NN | denotes | substrate |
| T1250 | 2428-2430 | VBZ | denotes | is |
| T1251 | 2431-2436 | NNP | denotes | HSP27 |
| T1252 | 2437-2438 | NNP | denotes | [ |
| T1253 | 2438-2440 | CD | denotes | 24 |
| T1254 | 2440-2441 | NNP | denotes | ] |
| T1255 | 2441-2442 | , | denotes | , |
| T1256 | 2443-2444 | DT | denotes | a |
| T1257 | 2445-2450 | JJ | denotes | small |
| T1258 | 2451-2455 | NN | denotes | heat |
| T1259 | 2456-2461 | NN | denotes | shock |
| T1260 | 2462-2469 | NN | denotes | protein |
| T1261 | 2470-2478 | VBN | denotes | involved |
| T1262 | 2479-2481 | IN | denotes | in |
| T1263 | 2482-2492 | VBG | denotes | regulating |
| T1264 | 2493-2497 | NN | denotes | cell |
| T1265 | 2498-2507 | NN | denotes | migration |
| T1266 | 2508-2511 | CC | denotes | and |
| T1267 | 2512-2516 | NN | denotes | cell |
| T1268 | 2517-2525 | NN | denotes | survival |
| T1269 | 2526-2527 | NNP | denotes | [ |
| T1270 | 2527-2529 | CD | denotes | 25 |
| T1271 | 2529-2530 | NNP | denotes | ] |
| T1272 | 2530-2531 | . | denotes | . |
| T1273 | 2532-2534 | DT | denotes | An |
| T1274 | 2535-2544 | JJ | denotes | essential |
| T1275 | 2545-2549 | NN | denotes | role |
| T1276 | 2550-2553 | IN | denotes | for |
| T1277 | 2554-2557 | NNP | denotes | PKD |
| T1278 | 2558-2565 | VBZ | denotes | enzymes |
| T1279 | 2566-2568 | IN | denotes | in |
| T1280 | 2569-2579 | VBG | denotes | regulating |
| T1281 | 2580-2585 | NN | denotes | class |
| T1282 | 2586-2588 | NNP | denotes | II |
| T1283 | 2589-2596 | NN | denotes | histone |
| T1284 | 2597-2609 | NNS | denotes | deacetylases |
| T1285 | 2610-2611 | -LRB- | denotes | ( |
| T1286 | 2611-2616 | NNS | denotes | HDACs |
| T1287 | 2616-2617 | -RRB- | denotes | ) |
| T1288 | 2617-2618 | , | denotes | , |
| T1289 | 2619-2626 | VBZ | denotes | enzymes |
| T1290 | 2627-2631 | IN | denotes | that |
| T1291 | 2632-2639 | VBZ | denotes | repress |
| T1292 | 2640-2654 | JJ | denotes | MEF2-dependent |
| T1293 | 2655-2659 | NN | denotes | gene |
| T1294 | 2660-2673 | NN | denotes | transcription |
| T1295 | 2673-2674 | , | denotes | , |
| T1296 | 2675-2678 | VBZ | denotes | has |
| T1297 | 2679-2683 | RB | denotes | also |
| T1298 | 2684-2688 | VBN | denotes | been |
| T1299 | 2689-2701 | VBN | denotes | demonstrated |
| T1300 | 2702-2703 | NNP | denotes | [ |
| T1301 | 2703-2707 | CD | denotes | 1,26 |
| T1302 | 2708-2710 | CD | denotes | 28 |
| T1303 | 2710-2711 | NN | denotes | ] |
| T1304 | 2711-2712 | . | denotes | . |
| T1305 | 2713-2715 | TO | denotes | To |
| T1306 | 2716-2727 | VB | denotes | investigate |
| T1307 | 2728-2731 | DT | denotes | the |
| T1308 | 2732-2742 | JJ | denotes | biological |
| T1309 | 2743-2747 | NN | denotes | role |
| T1310 | 2748-2750 | IN | denotes | of |
| T1311 | 2751-2755 | NNS | denotes | PKDs |
| T1312 | 2756-2758 | PRP | denotes | we |
| T1313 | 2759-2763 | VBP | denotes | have |
| T1314 | 2764-2773 | VBN | denotes | generated |
| T1315 | 2774-2778 | NNP | denotes | DT40 |
| T1316 | 2779-2780 | NNP | denotes | B |
| T1317 | 2781-2785 | NN | denotes | cell |
| T1318 | 2786-2791 | NNS | denotes | lines |
| T1319 | 2792-2796 | WDT | denotes | that |
| T1320 | 2797-2801 | VBP | denotes | lack |
| T1321 | 2802-2812 | NN | denotes | expression |
| T1322 | 2813-2815 | IN | denotes | of |
| T1323 | 2816-2819 | CD | denotes | one |
| T1324 | 2820-2822 | CC | denotes | or |
| T1325 | 2823-2827 | JJR | denotes | more |
| T1326 | 2828-2835 | NNS | denotes | members |
| T1327 | 2836-2838 | IN | denotes | of |
| T1328 | 2839-2842 | DT | denotes | the |
| T1329 | 2843-2846 | NNP | denotes | PKD |
| T1330 | 2847-2853 | NN | denotes | family |
| T1331 | 2854-2855 | NNP | denotes | [ |
| T1332 | 2855-2856 | CD | denotes | 1 |
| T1333 | 2856-2857 | NNP | denotes | ] |
| T1334 | 2857-2858 | , | denotes | , |
| T1335 | 2859-2867 | VBG | denotes | allowing |
| T1336 | 2868-2870 | PRP | denotes | us |
| T1337 | 2871-2873 | TO | denotes | to |
| T1338 | 2874-2885 | VB | denotes | investigate |
| T1339 | 2886-2889 | DT | denotes | the |
| T1340 | 2890-2898 | NN | denotes | function |
| T1341 | 2898-2899 | -LRB- | denotes | ( |
| T1342 | 2899-2900 | PRP | denotes | s |
| T1343 | 2900-2901 | -RRB- | denotes | ) |
| T1344 | 2902-2904 | IN | denotes | of |
| T1345 | 2905-2908 | NNP | denotes | PKD |
| T1346 | 2909-2917 | VBZ | denotes | isoforms |
| T1347 | 2918-2927 | VBG | denotes | following |
| T1348 | 2928-2929 | NNP | denotes | B |
| T1349 | 2930-2934 | NN | denotes | cell |
| T1350 | 2935-2942 | NN | denotes | antigen |
| T1351 | 2943-2951 | NN | denotes | receptor |
| T1352 | 2952-2953 | -LRB- | denotes | ( |
| T1353 | 2953-2956 | NNP | denotes | BCR |
| T1354 | 2956-2957 | -RRB- | denotes | ) |
| T1355 | 2958-2969 | NN | denotes | stimulation |
| T1356 | 2969-2970 | , | denotes | , |
| T1357 | 2971-2973 | RB | denotes | as |
| T1358 | 2974-2978 | RB | denotes | well |
| T1359 | 2979-2989 | VBG | denotes | addressing |
| T1360 | 2990-2993 | DT | denotes | the |
| T1361 | 2994-2999 | NN | denotes | issue |
| T1362 | 3000-3002 | IN | denotes | of |
| T1363 | 3003-3013 | JJ | denotes | functional |
| T1364 | 3014-3024 | NN | denotes | redundancy |
| T1365 | 3025-3032 | IN | denotes | between |
| T1366 | 3033-3036 | DT | denotes | the |
| T1367 | 3037-3046 | JJ | denotes | different |
| T1368 | 3047-3050 | NNP | denotes | PKD |
| T1369 | 3051-3057 | NN | denotes | family |
| T1370 | 3058-3065 | NNS | denotes | members |
| T1371 | 3065-3066 | . | denotes | . |
| T1372 | 3067-3075 | JJ | denotes | Previous |
| T1373 | 3076-3083 | NNS | denotes | studies |
| T1374 | 3084-3088 | VBP | denotes | have |
| T1375 | 3089-3094 | VBN | denotes | shown |
| T1376 | 3095-3099 | IN | denotes | that |
| T1377 | 3100-3104 | NNS | denotes | PKDs |
| T1378 | 3105-3108 | VBP | denotes | are |
| T1379 | 3109-3122 | JJ | denotes | indispensable |
| T1380 | 3123-3126 | IN | denotes | for |
| T1381 | 3127-3131 | NNP | denotes | HDAC |
| T1382 | 3132-3142 | NN | denotes | regulation |
| T1383 | 3143-3145 | IN | denotes | in |
| T1384 | 3146-3147 | NNP | denotes | B |
| T1385 | 3148-3153 | NNS | denotes | cells |
| T1386 | 3154-3155 | NNP | denotes | [ |
| T1387 | 3155-3156 | CD | denotes | 1 |
| T1388 | 3156-3157 | NNP | denotes | ] |
| T1389 | 3157-3158 | . | denotes | . |
| T1390 | 3159-3165 | NNP | denotes | Herein |
| T1391 | 3166-3168 | PRP | denotes | we |
| T1392 | 3169-3173 | VBP | denotes | show |
| T1393 | 3174-3178 | IN | denotes | that |
| T1394 | 3179-3183 | NNS | denotes | PKDs |
| T1395 | 3184-3187 | VBP | denotes | are |
| T1396 | 3188-3192 | RB | denotes | also |
| T1397 | 3193-3206 | JJ | denotes | indispensable |
| T1398 | 3207-3210 | IN | denotes | for |
| T1399 | 3211-3216 | NNP | denotes | HSP27 |
| T1400 | 3217-3232 | NN | denotes | phosphorylation |
| T1401 | 3233-3235 | IN | denotes | in |
| T1402 | 3236-3237 | NNP | denotes | B |
| T1403 | 3238-3243 | NNS | denotes | cells |
| T1404 | 3243-3244 | . | denotes | . |
| T1405 | 3245-3252 | RB | denotes | However |
| T1406 | 3252-3253 | , | denotes | , |
| T1407 | 3254-3262 | NNP | denotes | PKD-null |
| T1408 | 3263-3267 | NNP | denotes | DT40 |
| T1409 | 3268-3269 | NNP | denotes | B |
| T1410 | 3270-3275 | NNS | denotes | cells |
| T1411 | 3276-3279 | VBP | denotes | are |
| T1412 | 3280-3286 | JJ | denotes | viable |
| T1413 | 3287-3290 | CC | denotes | and |
| T1414 | 3291-3302 | VB | denotes | proliferate |
| T1415 | 3303-3311 | RB | denotes | normally |
| T1416 | 3311-3312 | . | denotes | . |
| T1417 | 3313-3321 | RB | denotes | Moreover |
| T1418 | 3321-3322 | , | denotes | , |
| T1419 | 3323-3327 | NN | denotes | loss |
| T1420 | 3328-3330 | IN | denotes | of |
| T1421 | 3331-3334 | DT | denotes | the |
| T1422 | 3335-3341 | JJ | denotes | entire |
| T1423 | 3342-3350 | JJ | denotes | cellular |
| T1424 | 3351-3355 | NN | denotes | pool |
| T1425 | 3356-3358 | IN | denotes | of |
| T1426 | 3359-3362 | NNP | denotes | PKD |
| T1427 | 3363-3367 | VBZ | denotes | does |
| T1428 | 3368-3371 | RB | denotes | not |
| T1429 | 3372-3382 | RB | denotes | critically |
| T1430 | 3383-3389 | VB | denotes | affect |
| T1431 | 3390-3399 | JJ | denotes | oxidative |
| T1432 | 3400-3406 | NN | denotes | stress |
| T1433 | 3407-3416 | NNS | denotes | responses |
| T1434 | 3417-3419 | IN | denotes | in |
| T1435 | 3420-3421 | NNP | denotes | B |
| T1436 | 3422-3427 | NNS | denotes | cells |
| T1437 | 3428-3431 | CC | denotes | nor |
| T1438 | 3432-3434 | VBP | denotes | do |
| T1439 | 3435-3438 | NNP | denotes | PKD |
| T1440 | 3439-3446 | VBZ | denotes | kinases |
| T1441 | 3447-3451 | VB | denotes | play |
| T1442 | 3452-3454 | DT | denotes | an |
| T1443 | 3455-3464 | JJ | denotes | essential |
| T1444 | 3465-3469 | NN | denotes | role |
| T1445 | 3470-3472 | IN | denotes | in |
| T1446 | 3473-3483 | VBG | denotes | regulating |
| T1447 | 3484-3488 | NNP | denotes | NFκB |
| T1448 | 3489-3504 | JJ | denotes | transcriptional |
| T1449 | 3505-3513 | NN | denotes | activity |
| T1450 | 3513-3514 | . | denotes | . |
| T1451 | 3515-3523 | RB | denotes | Together |
| T1452 | 3523-3524 | , | denotes | , |
| T1453 | 3525-3530 | DT | denotes | these |
| T1454 | 3531-3539 | NNS | denotes | findings |
| T1455 | 3540-3546 | VBP | denotes | reveal |
| T1456 | 3547-3551 | IN | denotes | that |
| T1457 | 3552-3554 | IN | denotes | in |
| T1458 | 3555-3556 | NNP | denotes | B |
| T1459 | 3557-3568 | NNS | denotes | lymphocytes |
| T1460 | 3568-3569 | , | denotes | , |
| T1461 | 3570-3573 | NNP | denotes | PKD |
| T1462 | 3574-3581 | NNS | denotes | kinases |
| T1463 | 3582-3585 | VBP | denotes | are |
| T1464 | 3586-3589 | RB | denotes | not |
| T1465 | 3590-3598 | JJ | denotes | critical |
| T1466 | 3599-3609 | NNS | denotes | regulators |
| T1467 | 3610-3612 | IN | denotes | of |
| T1468 | 3613-3617 | JJ | denotes | many |
| T1469 | 3618-3620 | IN | denotes | of |
| T1470 | 3621-3624 | DT | denotes | the |
| T1471 | 3625-3633 | JJ | denotes | cellular |
| T1472 | 3634-3643 | NNS | denotes | processes |
| T1473 | 3644-3654 | RB | denotes | previously |
| T1474 | 3655-3663 | VBD | denotes | ascribed |
| T1475 | 3664-3666 | TO | denotes | to |
| T1476 | 3667-3671 | PRP | denotes | them |
| T1477 | 3672-3674 | IN | denotes | in |
| T1478 | 3675-3680 | JJ | denotes | other |
| T1479 | 3681-3689 | JJ | denotes | cellular |
| T1480 | 3690-3697 | NNS | denotes | systems |
| T1481 | 3697-3698 | . | denotes | . |
| T1742 | 0-3731 | CD | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 |
| T1743 | 3731-3735 | NNP | denotes | Cell |
| T1744 | 3736-3743 | NN | denotes | culture |
| T1745 | 3743-3744 | , | denotes | , |
| T1746 | 3745-3754 | NN | denotes | transient |
| T1747 | 3755-3768 | NNS | denotes | transfections |
| T1748 | 3769-3772 | CC | denotes | and |
| T1749 | 3773-3777 | NN | denotes | cell |
| T1750 | 3778-3789 | NN | denotes | stimulation |
| T1751 | 3790-3793 | DT | denotes | The |
| T1752 | 3794-3804 | NN | denotes | generation |
| T1753 | 3804-3805 | , | denotes | , |
| T1754 | 3806-3813 | NN | denotes | culture |
| T1755 | 3814-3817 | CC | denotes | and |
| T1756 | 3818-3828 | NN | denotes | activation |
| T1757 | 3829-3831 | IN | denotes | of |
| T1758 | 3832-3836 | CD | denotes | PKD1 |
| T1759 | 3836-3837 | CD | denotes | − |
| T1760 | 3837-3838 | NN | denotes | / |
| T1761 | 3838-3839 | NN | denotes | − |
| T1762 | 3839-3840 | , | denotes | , |
| T1763 | 3841-3845 | NNP | denotes | PKD3 |
| T1764 | 3845-3846 | NNP | denotes | − |
| T1765 | 3846-3847 | VBD | denotes | / |
| T1766 | 3847-3848 | CD | denotes | − |
| T1767 | 3849-3852 | CC | denotes | and |
| T1768 | 3853-3859 | CD | denotes | PKD1/3 |
| T1769 | 3859-3860 | CD | denotes | − |
| T1770 | 3860-3861 | NN | denotes | / |
| T1771 | 3861-3862 | NN | denotes | − |
| T1772 | 3863-3871 | NN | denotes | knockout |
| T1773 | 3872-3876 | NNP | denotes | DT40 |
| T1774 | 3877-3878 | NNP | denotes | B |
| T1775 | 3879-3883 | NN | denotes | cell |
| T1776 | 3884-3889 | NNS | denotes | lines |
| T1777 | 3890-3894 | VBP | denotes | have |
| T1778 | 3895-3899 | VBN | denotes | been |
| T1779 | 3900-3909 | VBN | denotes | described |
| T1780 | 3910-3920 | RB | denotes | previously |
| T1781 | 3921-3922 | NNP | denotes | [ |
| T1782 | 3922-3923 | CD | denotes | 1 |
| T1783 | 3923-3924 | NNP | denotes | ] |
| T1784 | 3924-3925 | . | denotes | . |
| T1785 | 3926-3931 | NNS | denotes | Cells |
| T1786 | 3932-3936 | VBD | denotes | were |
| T1787 | 3937-3942 | VBN | denotes | lysed |
| T1788 | 3943-3946 | CC | denotes | and |
| T1789 | 3947-3954 | NN | denotes | protein |
| T1790 | 3955-3963 | NNS | denotes | extracts |
| T1791 | 3964-3968 | VBD | denotes | were |
| T1792 | 3969-3977 | VBN | denotes | analysed |
| T1793 | 3978-3980 | IN | denotes | in |
| T1794 | 3981-3988 | JJ | denotes | Western |
| T1795 | 3989-3997 | VBG | denotes | blotting |
| T1796 | 3998-4009 | NNS | denotes | experiments |
| T1797 | 4010-4012 | IN | denotes | as |
| T1798 | 4013-4023 | RB | denotes | previously |
| T1799 | 4024-4033 | VBN | denotes | described |
| T1800 | 4034-4035 | NNP | denotes | [ |
| T1801 | 4035-4036 | CD | denotes | 1 |
| T1802 | 4036-4037 | NNP | denotes | ] |
| T1803 | 4037-4038 | . | denotes | . |
| T1804 | 4039-4054 | NNP | denotes | Chloramphenicol |
| T1805 | 4055-4061 | NN | denotes | acetyl |
| T1806 | 4062-4073 | NN | denotes | transferase |
| T1807 | 4074-4080 | NNS | denotes | assays |
| T1808 | 4081-4085 | VBP | denotes | have |
| T1809 | 4086-4090 | VBN | denotes | been |
| T1810 | 4091-4100 | VBN | denotes | described |
| T1811 | 4101-4111 | RB | denotes | previously |
| T1812 | 4112-4113 | NNP | denotes | [ |
| T1813 | 4113-4115 | CD | denotes | 29 |
| T1814 | 4115-4116 | NNP | denotes | ] |
| T1815 | 4116-4117 | . | denotes | . |
| T1939 | 0-4124 | CD | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell culture, transient transfections and cell stimulation The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. Chloramphenicol acetyl transferase assays have been described previously [29]. 2.2 |
| T1940 | 4124-4128 | JJ | denotes | sIgM |
| T1941 | 4129-4137 | VBG | denotes | staining |
| R204 | T241 | T231 | dobj | enzymes,investigate |
| R205 | T242 | T243 | nsubj | we,generated |
| R206 | T243 | T241 | relcl | generated,enzymes |
| R207 | T244 | T249 | det | a,line |
| R208 | T245 | T249 | compound | PKD-null,line |
| R209 | T246 | T247 | compound | DT40,B-lymphocyte |
| R210 | T247 | T249 | compound | B-lymphocyte,line |
| R211 | T248 | T249 | compound | cell,line |
| R212 | T249 | T243 | dobj | line,generated |
| R213 | T250 | T216 | punct | .,are |
| R214 | T251 | T254 | advmod | Previously,shown |
| R215 | T252 | T254 | nsubj | we,shown |
| R216 | T253 | T254 | aux | have,shown |
| R217 | T254 | T254 | ROOT | shown,shown |
| R218 | T255 | T257 | mark | that,have |
| R219 | T256 | T257 | nsubj | PKDs,have |
| R220 | T257 | T254 | ccomp | have,shown |
| R221 | T258 | T260 | det | an,role |
| R222 | T259 | T260 | amod | essential,role |
| R223 | T260 | T257 | dobj | role,have |
| R224 | T261 | T260 | prep | in,role |
| R225 | T262 | T261 | pcomp | regulating,in |
| R226 | T263 | T266 | compound | class,deacetylases |
| R227 | T264 | T266 | compound | II,deacetylases |
| R228 | T265 | T266 | compound | histone,deacetylases |
| R229 | T266 | T262 | dobj | deacetylases,regulating |
| R230 | T267 | T262 | prep | in,regulating |
| R231 | T268 | T271 | compound | DT40,Matthews |
| R232 | T269 | T271 | compound | B-cells,Matthews |
| R233 | T270 | T271 | compound | [,Matthews |
| R234 | T271 | T267 | pobj | Matthews,in |
| R235 | T272 | T271 | punct | ",",Matthews |
| R236 | T273 | T271 | conj | S.A.,Matthews |
| R237 | T274 | T271 | punct | ",",Matthews |
| R238 | T275 | T271 | conj | Liu,Matthews |
| R239 | T276 | T275 | punct | ",",Liu |
| R240 | T277 | T275 | conj | P.,Liu |
| R241 | T278 | T277 | punct | ",",P. |
| R242 | T279 | T277 | conj | Spitaler,P. |
| R243 | T280 | T279 | punct | ",",Spitaler |
| R244 | T281 | T279 | conj | M.,Spitaler |
| R245 | T282 | T281 | punct | ",",M. |
| R246 | T283 | T281 | conj | Olson,M. |
| R247 | T284 | T283 | punct | ",",Olson |
| R248 | T285 | T283 | conj | E.N.,Olson |
| R249 | T286 | T285 | punct | ",",E.N. |
| R250 | T287 | T285 | conj | McKinsey,E.N. |
| R251 | T288 | T287 | punct | ",",McKinsey |
| R252 | T289 | T287 | conj | T.A.,McKinsey |
| R253 | T290 | T287 | punct | ",",McKinsey |
| R254 | T291 | T287 | conj | Cantrell,McKinsey |
| R255 | T292 | T291 | punct | ",",Cantrell |
| R256 | T293 | T291 | conj | D.A.,Cantrell |
| R257 | T294 | T293 | cc | and,D.A. |
| R258 | T295 | T293 | conj | Scharenberg,D.A. |
| R259 | T296 | T295 | punct | ",",Scharenberg |
| R260 | T297 | T295 | conj | A.M.,Scharenberg |
| R261 | T298 | T297 | punct | (,A.M. |
| R262 | T299 | T297 | appos | 2006,A.M. |
| R263 | T300 | T297 | punct | ),A.M. |
| R264 | T301 | T302 | amod | Essential,role |
| R265 | T302 | T260 | appos | role,role |
| R266 | T303 | T302 | prep | for,role |
| R267 | T304 | T303 | pobj | protein,for |
| R268 | T305 | T308 | compound | kinase,kinases |
| R269 | T306 | T307 | compound | D,family |
| R270 | T307 | T308 | compound | family,kinases |
| R271 | T308 | T302 | conj | kinases,role |
| R272 | T309 | T308 | prep | in,kinases |
| R273 | T310 | T311 | det | the,regulation |
| R274 | T311 | T309 | pobj | regulation,in |
| R275 | T312 | T311 | prep | of,regulation |
| R276 | T313 | T316 | compound | class,deacetylases |
| R277 | T314 | T316 | compound | II,deacetylases |
| R278 | T315 | T316 | compound | histone,deacetylases |
| R279 | T316 | T312 | pobj | deacetylases,of |
| R280 | T317 | T316 | prep | in,deacetylases |
| R281 | T318 | T319 | compound | B,lymphocytes |
| R282 | T319 | T317 | pobj | lymphocytes,in |
| R283 | T320 | T254 | punct | .,shown |
| R284 | T321 | T321 | ROOT | Mol,Mol |
| R285 | T322 | T321 | punct | .,Mol |
| R286 | T323 | T324 | compound | Cell,Biol |
| R287 | T324 | T324 | ROOT | Biol,Biol |
| R288 | T325 | T326 | punct | .,26 |
| R289 | T326 | T324 | nummod | 26,Biol |
| R290 | T327 | T324 | punct | ",",Biol |
| R291 | T328 | T324 | appos | 1569,Biol |
| R292 | T329 | T324 | appos | 1577,Biol |
| R293 | T330 | T330 | ROOT | ],] |
| R294 | T331 | T324 | punct | .,Biol |
| R295 | T332 | T334 | nsubj | We,show |
| R296 | T333 | T334 | advmod | now,show |
| R297 | T334 | T334 | ROOT | show,show |
| R298 | T335 | T339 | mark | that,required |
| R299 | T336 | T339 | nsubjpass | PKDs,required |
| R300 | T337 | T339 | auxpass | are,required |
| R301 | T338 | T339 | advmod | also,required |
| R302 | T339 | T334 | ccomp | required,show |
| R303 | T340 | T341 | aux | to,regulate |
| R304 | T341 | T339 | xcomp | regulate,required |
| R305 | T342 | T343 | compound | HSP27,phosphorylation |
| R306 | T343 | T341 | dobj | phosphorylation,regulate |
| R307 | T344 | T343 | prep | in,phosphorylation |
| R308 | T345 | T346 | compound | DT40,B-cells |
| R309 | T346 | T344 | pobj | B-cells,in |
| R310 | T347 | T334 | punct | .,show |
| R311 | T348 | T364 | advmod | However,regulate |
| R312 | T349 | T364 | punct | ",",regulate |
| R313 | T350 | T364 | prep | in,regulate |
| R314 | T351 | T350 | pobj | contrast,in |
| R315 | T352 | T351 | prep | to,contrast |
| R316 | T353 | T354 | amod | previous,observations |
| R317 | T354 | T352 | pobj | observations,to |
| R318 | T355 | T354 | prep | in,observations |
| R319 | T356 | T358 | amod | other,types |
| R320 | T357 | T358 | compound | cell,types |
| R321 | T358 | T355 | pobj | types,in |
| R322 | T359 | T364 | punct | ",",regulate |
| R323 | T360 | T364 | nsubj | PKD,regulate |
| R324 | T361 | T364 | nsubj | enzymes,regulate |
| R325 | T362 | T364 | aux | do,regulate |
| R326 | T363 | T364 | neg | not,regulate |
| R327 | T364 | T364 | ROOT | regulate,regulate |
| R328 | T365 | T367 | amod | basic,processes |
| R329 | T366 | T367 | amod | cellular,processes |
| R330 | T367 | T364 | dobj | processes,regulate |
| R331 | T368 | T369 | amod | such,as |
| R332 | T369 | T367 | prep | as,processes |
| R333 | T370 | T369 | pobj | proliferation,as |
| R334 | T371 | T370 | cc | or,proliferation |
| R335 | T372 | T370 | conj | survival,proliferation |
| R336 | T373 | T370 | conj | responses,proliferation |
| R337 | T374 | T373 | punct | ",",responses |
| R338 | T375 | T364 | cc | nor,regulate |
| R339 | T376 | T378 | nmod | NFκB,activity |
| R340 | T377 | T378 | amod | transcriptional,activity |
| R341 | T378 | T364 | conj | activity,regulate |
| R342 | T379 | T378 | amod | downstream,activity |
| R343 | T380 | T379 | prep | of,downstream |
| R344 | T381 | T385 | det | the,receptor |
| R345 | T382 | T384 | compound | B,antigen |
| R346 | T383 | T384 | compound | cell,antigen |
| R347 | T384 | T385 | compound | antigen,receptor |
| R348 | T385 | T380 | pobj | receptor,of |
| R349 | T386 | T364 | punct | .,regulate |
| R350 | T387 | T390 | advmod | Thus,have |
| R351 | T388 | T390 | punct | ",",have |
| R352 | T389 | T390 | nsubj | PKDs,have |
| R353 | T390 | T390 | ROOT | have,have |
| R354 | T391 | T393 | det | a,role |
| R355 | T392 | T393 | amod | selective,role |
| R356 | T393 | T390 | dobj | role,have |
| R357 | T394 | T393 | prep | in,role |
| R358 | T395 | T396 | compound | DT40,B-cell |
| R359 | T396 | T397 | compound | B-cell,biology |
| R360 | T397 | T394 | pobj | biology,in |
| R361 | T398 | T390 | punct | .,have |
| R894 | T1008 | T1011 | amod | "1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The",D |
| R895 | T1009 | T1011 | nmod | protein,D |
| R896 | T1010 | T1011 | compound | kinase,D |
| R897 | T1011 | T1011 | ROOT | D,D |
| R898 | T1012 | T1011 | punct | (,D |
| R899 | T1013 | T1011 | appos | PKD,D |
| R900 | T1014 | T1011 | punct | ),D |
| R901 | T1015 | T1017 | amod | serine/threonine,family |
| R902 | T1016 | T1017 | compound | kinase,family |
| R903 | T1017 | T1018 | nsubj | family,has |
| R904 | T1018 | T1018 | ROOT | has,has |
| R905 | T1019 | T1020 | nummod | three,members |
| R906 | T1020 | T1018 | dobj | members,has |
| R907 | T1021 | T1018 | punct | :,has |
| R908 | T1022 | T1022 | ROOT | PKD1,PKD1 |
| R909 | T1023 | T1022 | punct | ",",PKD1 |
| R910 | T1024 | T1022 | conj | PKD2,PKD1 |
| R911 | T1025 | T1024 | cc | and,PKD2 |
| R912 | T1026 | T1024 | conj | PKD3,PKD2 |
| R913 | T1027 | T1018 | punct | .,has |
| R914 | T1028 | T1030 | amod | Most,types |
| R915 | T1029 | T1030 | compound | cell,types |
| R916 | T1030 | T1031 | nsubj | types,express |
| R917 | T1031 | T1031 | ROOT | express,express |
| R918 | T1032 | T1033 | advmod | at,least |
| R919 | T1033 | T1034 | advmod | least,two |
| R920 | T1034 | T1036 | nummod | two,isoforms |
| R921 | T1035 | T1036 | compound | PKD,isoforms |
| R922 | T1036 | T1031 | dobj | isoforms,express |
| R923 | T1037 | T1031 | cc | but,express |
| R924 | T1038 | T1039 | compound | PKD,enzymes |
| R925 | T1039 | T1040 | nsubj | enzymes,are |
| R926 | T1040 | T1043 | auxpass | are,expressed |
| R927 | T1041 | T1042 | advmod | especially,highly |
| R928 | T1042 | T1043 | advmod | highly,expressed |
| R929 | T1043 | T1031 | conj | expressed,express |
| R930 | T1044 | T1043 | prep | in,expressed |
| R931 | T1045 | T1046 | amod | haematopoietic,cells |
| R932 | T1046 | T1044 | pobj | cells,in |
| R933 | T1047 | T1043 | punct | ",",expressed |
| R934 | T1048 | T1051 | advmod | where,activated |
| R935 | T1049 | T1051 | nsubjpass | they,activated |
| R936 | T1050 | T1051 | auxpass | are,activated |
| R937 | T1051 | T1043 | advcl | activated,expressed |
| R938 | T1052 | T1051 | prep | in,activated |
| R939 | T1053 | T1052 | pobj | response,in |
| R940 | T1054 | T1053 | prep | to,response |
| R941 | T1055 | T1056 | compound | antigen,receptors |
| R942 | T1056 | T1057 | compound | receptors,stimulation |
| R943 | T1057 | T1054 | pobj | stimulation,to |
| R944 | T1058 | T1060 | compound | [,] |
| R945 | T1059 | T1060 | compound | "2,3",] |
| R946 | T1060 | T1057 | appos | ],stimulation |
| R947 | T1061 | T1043 | punct | .,expressed |
| R948 | T1062 | T1065 | det | A,pathway |
| R949 | T1063 | T1065 | amod | conserved,pathway |
| R950 | T1064 | T1065 | amod | signalling,pathway |
| R951 | T1065 | T1071 | nsubj | pathway,involves |
| R952 | T1066 | T1065 | acl | linking,pathway |
| R953 | T1067 | T1068 | compound | antigen,receptors |
| R954 | T1068 | T1066 | dobj | receptors,linking |
| R955 | T1069 | T1066 | prep | to,linking |
| R956 | T1070 | T1069 | pobj | PKDs,to |
| R957 | T1071 | T1071 | ROOT | involves,involves |
| R958 | T1072 | T1073 | det | the,activation |
| R959 | T1073 | T1071 | dobj | activation,involves |
| R960 | T1074 | T1073 | prep | of,activation |
| R961 | T1075 | T1074 | pobj | PLCγ,of |
| R962 | T1076 | T1073 | cc | and,activation |
| R963 | T1077 | T1079 | det | the,production |
| R964 | T1078 | T1079 | amod | subsequent,production |
| R965 | T1079 | T1073 | conj | production,activation |
| R966 | T1080 | T1079 | prep | of,production |
| R967 | T1081 | T1080 | pobj | diacylglycerol,of |
| R968 | T1082 | T1083 | punct | (,DAG |
| R969 | T1083 | T1081 | appos | DAG,diacylglycerol |
| R970 | T1084 | T1081 | punct | ),diacylglycerol |
| R971 | T1085 | T1086 | nsubj | which,stimulates |
| R972 | T1086 | T1079 | relcl | stimulates,production |
| R973 | T1087 | T1090 | amod | classical,protein |
| R974 | T1088 | T1087 | cc | and/or,classical |
| R975 | T1089 | T1087 | conj | novel,classical |
| R976 | T1090 | T1092 | compound | protein,Cs |
| R977 | T1091 | T1092 | compound | kinase,Cs |
| R978 | T1092 | T1086 | dobj | Cs,stimulates |
| R979 | T1093 | T1092 | punct | (,Cs |
| R980 | T1094 | T1092 | appos | PKC,Cs |
| R981 | T1095 | T1092 | punct | ),Cs |
| R982 | T1096 | T1109 | nsubj | that,kinases |
| R983 | T1097 | T1096 | aux | phosphorylate,that |
| R984 | T1098 | T1102 | nummod | two,residues |
| R985 | T1099 | T1102 | amod | key,residues |
| R986 | T1100 | T1102 | amod | regulatory,residues |
| R987 | T1101 | T1102 | compound | serine,residues |
| R988 | T1102 | T1097 | dobj | residues,phosphorylate |
| R989 | T1103 | T1102 | prep | in,residues |
| R990 | T1104 | T1106 | det | the,loop |
| R991 | T1105 | T1106 | compound | activation,loop |
| R992 | T1106 | T1103 | pobj | loop,in |
| R993 | T1107 | T1106 | prep | of,loop |
| R994 | T1108 | T1107 | pobj | PKD,of |
| R995 | T1109 | T1079 | relcl | kinases,production |
| R996 | T1110 | T1109 | dobj | [,kinases |
| R997 | T1111 | T1110 | nummod | 3,[ |
| R998 | T1112 | T1113 | nummod | 6,] |
| R999 | T1113 | T1079 | appos | ],production |
| R1000 | T1114 | T1071 | punct | .,involves |
| R1001 | T1115 | T1118 | det | The,region |
| R1002 | T1116 | T1118 | amod | N-terminal,region |
| R1003 | T1117 | T1118 | amod | regulatory,region |
| R1004 | T1118 | T1121 | nsubj | region,enzymes |
| R1005 | T1119 | T1118 | prep | of,region |
| R1006 | T1120 | T1119 | pobj | PKD,of |
| R1007 | T1121 | T1121 | ROOT | enzymes,enzymes |
| R1008 | T1122 | T1121 | conj | contains,enzymes |
| R1009 | T1123 | T1124 | det | a,DAG |
| R1010 | T1124 | T1126 | nmod | DAG,domain |
| R1011 | T1125 | T1126 | amod | binding,domain |
| R1012 | T1126 | T1122 | dobj | domain,contains |
| R1013 | T1127 | T1126 | cc | and,domain |
| R1014 | T1128 | T1129 | amod | direct,binding |
| R1015 | T1129 | T1126 | conj | binding,domain |
| R1016 | T1130 | T1129 | prep | of,binding |
| R1017 | T1131 | T1130 | pobj | DAG,of |
| R1018 | T1132 | T1133 | advmod | also,contributes |
| R1019 | T1133 | T1121 | conj | contributes,enzymes |
| R1020 | T1134 | T1133 | prep | to,contributes |
| R1021 | T1135 | T1139 | nummod | PKD1,] |
| R1022 | T1136 | T1137 | compound | activation,[ |
| R1023 | T1137 | T1139 | nmod | [,] |
| R1024 | T1138 | T1137 | nummod | 7,[ |
| R1025 | T1139 | T1134 | pobj | ],to |
| R1026 | T1140 | T1142 | advmod | as,as |
| R1027 | T1141 | T1142 | advmod | well,as |
| R1028 | T1142 | T1133 | prep | as,contributes |
| R1029 | T1143 | T1142 | pcomp | regulating,as |
| R1030 | T1144 | T1146 | det | the,location |
| R1031 | T1145 | T1146 | amod | spatial,location |
| R1032 | T1146 | T1143 | dobj | location,regulating |
| R1033 | T1147 | T1146 | prep | of,location |
| R1034 | T1148 | T1149 | compound | PKD,enzymes |
| R1035 | T1149 | T1147 | pobj | enzymes,of |
| R1036 | T1150 | T1143 | prep | within,regulating |
| R1037 | T1151 | T1150 | pobj | cells,within |
| R1038 | T1152 | T1153 | nmod | [,8 |
| R1039 | T1153 | T1143 | npadvmod | 8,regulating |
| R1040 | T1154 | T1155 | nummod | 12,] |
| R1041 | T1155 | T1153 | appos | ],8 |
| R1042 | T1156 | T1121 | punct | .,enzymes |
| R1043 | T1157 | T1158 | compound | PKD,enzymes |
| R1044 | T1158 | T1161 | nsubjpass | enzymes,proposed |
| R1045 | T1159 | T1161 | aux | have,proposed |
| R1062 | T1176 | T1177 | amod | anti-apoptotic,signals |
| R1063 | T1177 | T1171 | appos | signals,[ |
| R1064 | T1178 | T1180 | compound | [,] |
| R1065 | T1179 | T1180 | compound | "17,18",] |
| R1066 | T1180 | T1177 | dobj | ],signals |
| R1067 | T1181 | T1180 | cc | and,] |
| R1068 | T1182 | T1183 | amod | thymocyte,development |
| R1069 | T1183 | T1186 | nmod | development,] |
| R1070 | T1184 | T1186 | nmod | [,] |
| R1071 | T1185 | T1186 | nummod | 19,] |
| R1072 | T1186 | T1180 | conj | ],] |
| R1073 | T1187 | T1177 | punct | .,signals |
| R1074 | T1188 | T1199 | nsubj | Expression,modify |
| R1075 | T1189 | T1188 | prep | of,Expression |
| R1076 | T1190 | T1192 | amod | mutant,inactive |
| R1077 | T1191 | T1192 | advmod | catalytically,inactive |
| R1078 | T1192 | T1196 | amod | inactive,PKDs |
| R1079 | T1193 | T1192 | cc | and,inactive |
| R1080 | T1194 | T1195 | advmod | constitutively,activated |
| R1081 | T1195 | T1192 | conj | activated,inactive |
| R1082 | T1196 | T1188 | appos | PKDs,Expression |
| R1083 | T1197 | T1199 | aux | can,modify |
| R1084 | T1198 | T1199 | advmod | also,modify |
| R1085 | T1199 | T1199 | ROOT | modify,modify |
| R1086 | T1200 | T1201 | compound | Golgi,function |
| R1087 | T1201 | T1199 | dobj | function,modify |
| R1088 | T1202 | T1201 | punct | ",",function |
| R1089 | T1203 | T1204 | compound | cell,adhesion |
| R1090 | T1204 | T1201 | conj | adhesion,function |
| R1091 | T1205 | T1204 | cc | and,adhesion |
| R1092 | T1206 | T1207 | compound | cell,motility |
| R1093 | T1207 | T1204 | conj | motility,adhesion |
| R1094 | T1208 | T1209 | punct | (,reviewed |
| R1095 | T1209 | T1199 | advcl | reviewed,modify |
| R1096 | T1210 | T1209 | prep | in,reviewed |
| R1097 | T1211 | T1210 | pobj | [,in |
| R1098 | T1212 | T1213 | nummod | 20,] |
| R1099 | T1213 | T1211 | appos | ],[ |
| R1100 | T1214 | T1211 | punct | ),[ |
| R1101 | T1215 | T1199 | punct | .,modify |
| R1102 | T1216 | T1223 | prep | In,linked |
| R1103 | T1217 | T1216 | amod | particular,In |
| R1104 | T1218 | T1223 | punct | ",",linked |
| R1105 | T1219 | T1223 | nsubjpass | PKDs,linked |
| R1106 | T1220 | T1223 | aux | have,linked |
| R1107 | T1221 | T1223 | auxpass | been,linked |
| R1108 | T1222 | T1223 | advmod | widely,linked |
| R1109 | T1223 | T1223 | ROOT | linked,linked |
| R1110 | T1224 | T1223 | prep | to,linked |
| R1111 | T1225 | T1226 | det | the,activation |
| R1112 | T1226 | T1224 | pobj | activation,to |
| R1113 | T1227 | T1226 | prep | of,activation |
| R1114 | T1228 | T1231 | det | the,factor |
| R1115 | T1229 | T1231 | compound | NFκB,factor |
| R1116 | T1230 | T1231 | compound | transcription,factor |
| R1117 | T1231 | T1227 | pobj | factor,of |
| R1118 | T1232 | T1224 | cc | and,to |
| R1119 | T1233 | T1223 | prep | in,linked |
| R1120 | T1234 | T1233 | pcomp | regulating,in |
| R1121 | T1235 | T1236 | compound | cell,survival |
| R1122 | T1236 | T1234 | dobj | survival,regulating |
| R1123 | T1237 | T1234 | prep | during,regulating |
| R1124 | T1238 | T1239 | amod | oxidative,stress |
| R1125 | T1239 | T1241 | compound | stress,"17,21" |
| R1126 | T1240 | T1241 | compound | [,"17,21" |
| R1127 | T1241 | T1237 | pobj | "17,21",during |
| R1175 | T1289 | T1289 | ROOT | enzymes,enzymes |
| R1176 | T1290 | T1291 | mark | that,repress |
| R1196 | T1310 | T1309 | prep | of,role |
| R1197 | T1311 | T1310 | pobj | PKDs,of |
| R1198 | T1312 | T1314 | nsubj | we,generated |
| R1199 | T1313 | T1314 | aux | have,generated |
| R1200 | T1314 | T1314 | ROOT | generated,generated |
| R1201 | T1315 | T1318 | compound | DT40,lines |
| R1202 | T1316 | T1318 | compound | B,lines |
| R1203 | T1317 | T1318 | compound | cell,lines |
| R1204 | T1318 | T1314 | dobj | lines,generated |
| R1205 | T1319 | T1320 | nsubj | that,lack |
| R1206 | T1320 | T1318 | relcl | lack,lines |
| R1207 | T1321 | T1320 | dobj | expression,lack |
| R1208 | T1322 | T1321 | prep | of,expression |
| R1209 | T1323 | T1326 | nummod | one,members |
| R1210 | T1324 | T1323 | cc | or,one |
| R1211 | T1325 | T1323 | conj | more,one |
| R1212 | T1326 | T1322 | pobj | members,of |
| R1213 | T1327 | T1326 | prep | of,members |
| R1214 | T1328 | T1330 | det | the,family |
| R1215 | T1329 | T1330 | compound | PKD,family |
| R1216 | T1330 | T1327 | pobj | family,of |
| R1217 | T1331 | T1333 | nmod | [,] |
| R1218 | T1332 | T1333 | nummod | 1,] |
| R1219 | T1333 | T1326 | appos | ],members |
| R1220 | T1334 | T1314 | punct | ",",generated |
| R1221 | T1335 | T1314 | advcl | allowing,generated |
| R1222 | T1336 | T1338 | nsubj | us,investigate |
| R1223 | T1337 | T1338 | aux | to,investigate |
| R1224 | T1338 | T1335 | ccomp | investigate,allowing |
| R1225 | T1339 | T1340 | det | the,function |
| R1226 | T1340 | T1338 | dobj | function,investigate |
| R1227 | T1341 | T1340 | punct | (,function |
| R1228 | T1342 | T1340 | appos | s,function |
| R1229 | T1343 | T1340 | punct | ),function |
| R1230 | T1344 | T1340 | prep | of,function |
| R1231 | T1345 | T1344 | pobj | PKD,of |
| R1232 | T1346 | T1344 | pobj | isoforms,of |
| R1233 | T1347 | T1346 | acl | following,isoforms |
| R1234 | T1348 | T1350 | compound | B,antigen |
| R1235 | T1349 | T1350 | compound | cell,antigen |
| R1236 | T1350 | T1351 | compound | antigen,receptor |
| R1237 | T1351 | T1347 | dobj | receptor,following |
| R1238 | T1352 | T1353 | punct | (,BCR |
| R1239 | T1353 | T1351 | appos | BCR,receptor |
| R1240 | T1354 | T1353 | punct | ),BCR |
| R1241 | T1355 | T1351 | conj | stimulation,receptor |
| R1242 | T1356 | T1347 | punct | ",",following |
| R1243 | T1357 | T1359 | advmod | as,addressing |
| R1244 | T1358 | T1359 | advmod | well,addressing |
| R1245 | T1359 | T1338 | advcl | addressing,investigate |
| R1246 | T1360 | T1361 | det | the,issue |
| R1247 | T1361 | T1359 | dobj | issue,addressing |
| R1248 | T1362 | T1361 | prep | of,issue |
| R1249 | T1363 | T1364 | amod | functional,redundancy |
| R1250 | T1364 | T1362 | pobj | redundancy,of |
| R1251 | T1365 | T1364 | prep | between,redundancy |
| R1252 | T1366 | T1370 | det | the,members |
| R1253 | T1367 | T1370 | amod | different,members |
| R1254 | T1368 | T1370 | compound | PKD,members |
| R1255 | T1369 | T1370 | compound | family,members |
| R1256 | T1370 | T1365 | pobj | members,between |
| R1257 | T1371 | T1314 | punct | .,generated |
| R1258 | T1372 | T1373 | amod | Previous,studies |
| R1259 | T1373 | T1375 | nsubj | studies,shown |
| R1261 | T1374 | T1375 | aux | have,shown |
| R1264 | T1375 | T1375 | ROOT | shown,shown |
| R1267 | T1376 | T1378 | mark | that,are |
| R1275 | T1377 | T1378 | nsubj | PKDs,are |
| R1279 | T1378 | T1375 | ccomp | are,shown |
| R1284 | T1379 | T1378 | acomp | indispensable,are |
| R1288 | T1380 | T1378 | prep | for,are |
| R1292 | T1381 | T1382 | compound | HDAC,regulation |
| R1296 | T1382 | T1380 | pobj | regulation,for |
| R1298 | T1383 | T1382 | prep | in,regulation |
| R1299 | T1477 | T1474 | prep | in,ascribed |
| R1300 | T1478 | T1480 | amod | other,systems |
| R1301 | T1479 | T1480 | amod | cellular,systems |
| R1302 | T1480 | T1477 | pobj | systems,in |
| R1303 | T1384 | T1385 | compound | B,cells |
| R1304 | T1481 | T1455 | punct | .,reveal |
| R1305 | T1385 | T1383 | pobj | cells,in |
| R1306 | T1386 | T1388 | nmod | [,] |
| R1307 | T1387 | T1388 | nummod | 1,] |
| R1308 | T1388 | T1382 | appos | ],regulation |
| R1309 | T1389 | T1375 | punct | .,shown |
| R1310 | T1390 | T1392 | npadvmod | Herein,show |
| R1311 | T1391 | T1392 | nsubj | we,show |
| R1312 | T1392 | T1392 | ROOT | show,show |
| R1313 | T1393 | T1395 | mark | that,are |
| R1314 | T1394 | T1395 | nsubj | PKDs,are |
| R1315 | T1395 | T1392 | ccomp | are,show |
| R1316 | T1396 | T1395 | advmod | also,are |
| R1317 | T1397 | T1395 | acomp | indispensable,are |
| R1318 | T1398 | T1395 | prep | for,are |
| R1319 | T1399 | T1400 | compound | HSP27,phosphorylation |
| R1320 | T1400 | T1398 | pobj | phosphorylation,for |
| R1321 | T1401 | T1400 | prep | in,phosphorylation |
| R1322 | T1402 | T1403 | compound | B,cells |
| R1323 | T1403 | T1401 | pobj | cells,in |
| R1324 | T1404 | T1392 | punct | .,show |
| R1325 | T1405 | T1411 | advmod | However,are |
| R1326 | T1406 | T1411 | punct | ",",are |
| R1327 | T1407 | T1409 | compound | PKD-null,B |
| R1328 | T1408 | T1409 | compound | DT40,B |
| R1329 | T1409 | T1410 | compound | B,cells |
| R1331 | T1410 | T1411 | nsubj | cells,are |
| R1335 | T1411 | T1411 | ROOT | are,are |
| R1336 | T1412 | T1411 | acomp | viable,are |
| R1338 | T1413 | T1411 | cc | and,are |
| R1340 | T1414 | T1411 | conj | proliferate,are |
| R1341 | T1415 | T1414 | advmod | normally,proliferate |
| R1342 | T1416 | T1411 | punct | .,are |
| R1343 | T1417 | T1430 | advmod | Moreover,affect |
| R1344 | T1418 | T1430 | punct | ",",affect |
| R1345 | T1419 | T1430 | nsubj | loss,affect |
| R1347 | T1420 | T1419 | prep | of,loss |
| R1351 | T1421 | T1424 | det | the,pool |
| R1352 | T1422 | T1424 | amod | entire,pool |
| R1355 | T1423 | T1424 | amod | cellular,pool |
| R1356 | T1424 | T1420 | pobj | pool,of |
| R1357 | T1425 | T1424 | prep | of,pool |
| R1358 | T1426 | T1425 | pobj | PKD,of |
| R1359 | T1427 | T1430 | aux | does,affect |
| R1360 | T1428 | T1430 | neg | not,affect |
| R1361 | T1429 | T1430 | advmod | critically,affect |
| R1362 | T1430 | T1430 | ROOT | affect,affect |
| R1365 | T1431 | T1433 | amod | oxidative,responses |
| R1367 | T1432 | T1433 | compound | stress,responses |
| R1369 | T1433 | T1430 | dobj | responses,affect |
| R1371 | T1434 | T1433 | prep | in,responses |
| R1373 | T1435 | T1436 | compound | B,cells |
| R1374 | T1436 | T1434 | pobj | cells,in |
| R1376 | T1437 | T1430 | cc | nor,affect |
| R1378 | T1438 | T1441 | aux | do,play |
| R1379 | T1439 | T1440 | compound | PKD,kinases |
| R1380 | T1440 | T1441 | nsubj | kinases,play |
| R1381 | T1441 | T1430 | conj | play,affect |
| R1382 | T1442 | T1444 | det | an,role |
| R1383 | T1443 | T1444 | amod | essential,role |
| R1384 | T1444 | T1441 | dobj | role,play |
| R1385 | T1445 | T1444 | prep | in,role |
| R1387 | T1446 | T1445 | pcomp | regulating,in |
| R1391 | T1447 | T1449 | nmod | NFκB,activity |
| R1519 | T1759 | T1761 | nummod | −,− |
| R1520 | T1760 | T1761 | compound | /,− |
| R1521 | T1761 | T1757 | pobj | −,of |
| R1522 | T1762 | T1765 | punct | ",",/ |
| R1523 | T1763 | T1764 | compound | PKD3,− |
| R1524 | T1764 | T1765 | nsubj | −,/ |
| R1525 | T1765 | T1765 | ROOT | /,/ |
| R1526 | T1766 | T1776 | nummod | −,lines |
| R1527 | T1767 | T1766 | cc | and,− |
| R1528 | T1768 | T1776 | nummod | PKD1/3,lines |
| R1529 | T1769 | T1776 | nummod | −,lines |
| R1530 | T1770 | T1772 | nmod | /,knockout |
| R1531 | T1771 | T1772 | compound | −,knockout |
| R1532 | T1772 | T1776 | compound | knockout,lines |
| R1533 | T1773 | T1774 | compound | DT40,B |
| R1534 | T1774 | T1775 | compound | B,cell |
| R1535 | T1775 | T1776 | compound | cell,lines |
| R1536 | T1776 | T1779 | nsubjpass | lines,described |
| R1537 | T1777 | T1779 | aux | have,described |
| R1538 | T1778 | T1779 | auxpass | been,described |
| R1539 | T1779 | T1765 | ccomp | described,/ |
| R1540 | T1780 | T1783 | advmod | previously,] |
| R1541 | T1781 | T1783 | nmod | [,] |
| R1542 | T1782 | T1781 | nummod | 1,[ |
| R1543 | T1783 | T1779 | dobj | ],described |
| R1544 | T1784 | T1765 | punct | .,/ |
| R1545 | T1785 | T1787 | nsubjpass | Cells,lysed |
| R1546 | T1786 | T1787 | auxpass | were,lysed |
| R1547 | T1787 | T1787 | ROOT | lysed,lysed |
| R1548 | T1788 | T1787 | cc | and,lysed |
| R1549 | T1789 | T1790 | compound | protein,extracts |
| R1550 | T1790 | T1792 | nsubjpass | extracts,analysed |
| R1551 | T1791 | T1792 | auxpass | were,analysed |
| R1552 | T1792 | T1787 | conj | analysed,lysed |
| R1553 | T1793 | T1792 | prep | in,analysed |
| R1554 | T1794 | T1796 | amod | Western,experiments |
| R1555 | T1795 | T1796 | amod | blotting,experiments |
| R1556 | T1796 | T1793 | pobj | experiments,in |
| R1557 | T1797 | T1799 | mark | as,described |
| R1558 | T1798 | T1799 | advmod | previously,described |
| R1559 | T1799 | T1792 | advcl | described,analysed |
| R1560 | T1800 | T1802 | nmod | [,] |
| R1561 | T1801 | T1802 | nummod | 1,] |
| R1562 | T1802 | T1799 | dobj | ],described |
| R1563 | T1803 | T1792 | punct | .,analysed |
| R1564 | T1804 | T1807 | nmod | Chloramphenicol,assays |
| R1565 | T1805 | T1807 | nmod | acetyl,assays |
| R1566 | T1806 | T1807 | compound | transferase,assays |
| R1567 | T1807 | T1810 | nsubjpass | assays,described |
| R1568 | T1808 | T1810 | aux | have,described |
| R1569 | T1809 | T1810 | auxpass | been,described |
| R1570 | T1810 | T1810 | ROOT | described,described |
| R1571 | T1811 | T1814 | advmod | previously,] |
| R1572 | T1812 | T1814 | nmod | [,] |
| R1573 | T1813 | T1814 | nummod | 29,] |
| R1574 | T1814 | T1810 | dobj | ],described |
| R1575 | T1815 | T1810 | punct | .,described |
| R1663 | T1939 | T1940 | nummod | "Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell culture, transient transfections and cell stimulation The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. Chloramphenicol acetyl transferase assays have been described previously [29]. 2.2 ",sIgM |
| R1050 | T1164 | T1166 | amod | numerous,functions |
| R1051 | T1165 | T1166 | amod | cellular,functions |
| R1052 | T1166 | T1163 | dobj | functions,regulate |
| R1053 | T1167 | T1166 | punct | ",",functions |
| R1054 | T1168 | T1166 | prep | including,functions |
| R1055 | T1169 | T1170 | compound | cell,proliferation |
| R1056 | T1170 | T1168 | pobj | proliferation,including |
| R1057 | T1171 | T1170 | dobj | [,proliferation |
| R1058 | T1172 | T1171 | nummod | 13,[ |
| R1059 | T1173 | T1174 | nummod | 16,] |
| R1060 | T1174 | T1171 | appos | ],[ |
| R1061 | T1175 | T1177 | punct | ",",signals |
| R1131 | T1245 | T1249 | det | Another,substrate |
| R1132 | T1246 | T1247 | advmod | recently,proposed |
| R1133 | T1247 | T1249 | amod | proposed,substrate |
| R1134 | T1248 | T1249 | compound | PKD1,substrate |
| R1135 | T1249 | T1250 | nsubj | substrate,is |
| R1136 | T1250 | T1250 | ROOT | is,is |
| R1137 | T1251 | T1252 | compound | HSP27,[ |
| R1138 | T1252 | T1250 | attr | [,is |
| R1139 | T1253 | T1254 | nummod | 24,] |
| R1140 | T1254 | T1252 | appos | ],[ |
| R1141 | T1255 | T1254 | punct | ",",] |
| R1142 | T1256 | T1260 | det | a,protein |
| R1143 | T1257 | T1260 | amod | small,protein |
| R1144 | T1258 | T1259 | compound | heat,shock |
| R1145 | T1259 | T1260 | compound | shock,protein |
| R1146 | T1260 | T1254 | appos | protein,] |
| R1147 | T1261 | T1260 | amod | involved,protein |
| R1148 | T1262 | T1261 | prep | in,involved |
| R1149 | T1263 | T1262 | pcomp | regulating,in |
| R1150 | T1264 | T1265 | compound | cell,migration |
| R1151 | T1265 | T1263 | dobj | migration,regulating |
| R1152 | T1266 | T1265 | cc | and,migration |
| R1153 | T1267 | T1268 | compound | cell,survival |
| R1154 | T1268 | T1265 | conj | survival,migration |
| R1155 | T1269 | T1271 | nmod | [,] |
| R1156 | T1270 | T1271 | nummod | 25,] |
| R1157 | T1271 | T1265 | appos | ],migration |
| R1158 | T1272 | T1250 | punct | .,is |
| R1159 | T1273 | T1275 | det | An,role |
| R1160 | T1274 | T1275 | amod | essential,role |
| R1161 | T1275 | T1289 | nsubj | role,enzymes |
| R1162 | T1276 | T1275 | prep | for,role |
| R1163 | T1277 | T1278 | compound | PKD,enzymes |
| R1164 | T1278 | T1276 | pobj | enzymes,for |
| R1165 | T1279 | T1278 | prep | in,enzymes |
| R1166 | T1280 | T1279 | pcomp | regulating,in |
| R1167 | T1281 | T1284 | nmod | class,deacetylases |
| R1168 | T1282 | T1281 | nummod | II,class |
| R1169 | T1283 | T1284 | compound | histone,deacetylases |
| R1170 | T1284 | T1280 | dobj | deacetylases,regulating |
| R1171 | T1285 | T1284 | punct | (,deacetylases |
| R1172 | T1286 | T1284 | appos | HDACs,deacetylases |
| R1173 | T1287 | T1284 | punct | ),deacetylases |
| R1174 | T1288 | T1278 | punct | ",",enzymes |
| R1177 | T1291 | T1289 | ccomp | repress,enzymes |
| R1178 | T1292 | T1294 | amod | MEF2-dependent,transcription |
| R1179 | T1293 | T1294 | compound | gene,transcription |
| R1180 | T1294 | T1291 | dobj | transcription,repress |
| R1181 | T1295 | T1299 | punct | ",",demonstrated |
| R1182 | T1296 | T1299 | aux | has,demonstrated |
| R1183 | T1297 | T1299 | advmod | also,demonstrated |
| R1184 | T1298 | T1299 | auxpass | been,demonstrated |
| R1185 | T1299 | T1289 | conj | demonstrated,enzymes |
| R1186 | T1300 | T1301 | compound | [,"1,26" |
| R1187 | T1301 | T1299 | dobj | "1,26",demonstrated |
| R1188 | T1302 | T1303 | nummod | 28,] |
| R1189 | T1303 | T1299 | npadvmod | ],demonstrated |
| R1190 | T1304 | T1299 | punct | .,demonstrated |
| R1191 | T1305 | T1306 | aux | To,investigate |
| R1192 | T1306 | T1314 | advcl | investigate,generated |
| R1193 | T1307 | T1309 | det | the,role |
| R1194 | T1308 | T1309 | amod | biological,role |
| R1195 | T1309 | T1306 | dobj | role,investigate |
| R1260 | T1448 | T1449 | amod | transcriptional,activity |
| R1262 | T1449 | T1446 | dobj | activity,regulating |
| R1263 | T1450 | T1441 | punct | .,play |
| R1265 | T1451 | T1455 | advmod | Together,reveal |
| R1266 | T1452 | T1455 | punct | ",",reveal |
| R1268 | T1453 | T1454 | det | these,findings |
| R1269 | T1454 | T1455 | nsubj | findings,reveal |
| R1270 | T1455 | T1455 | ROOT | reveal,reveal |
| R1271 | T1456 | T1463 | mark | that,are |
| R1272 | T1457 | T1463 | prep | in,are |
| R1273 | T1458 | T1459 | compound | B,lymphocytes |
| R1274 | T1459 | T1457 | pobj | lymphocytes,in |
| R1276 | T1460 | T1463 | punct | ",",are |
| R1277 | T1461 | T1462 | compound | PKD,kinases |
| R1278 | T1462 | T1463 | nsubj | kinases,are |
| R1280 | T1463 | T1455 | ccomp | are,reveal |
| R1281 | T1464 | T1463 | neg | not,are |
| R1282 | T1465 | T1466 | amod | critical,regulators |
| R1283 | T1466 | T1463 | attr | regulators,are |
| R1285 | T1467 | T1466 | prep | of,regulators |
| R1286 | T1468 | T1467 | pobj | many,of |
| R1287 | T1469 | T1468 | prep | of,many |
| R1289 | T1470 | T1472 | det | the,processes |
| R1290 | T1471 | T1472 | amod | cellular,processes |
| R1291 | T1472 | T1469 | pobj | processes,of |
| R1293 | T1473 | T1474 | advmod | previously,ascribed |
| R1294 | T1474 | T1463 | conj | ascribed,are |
| R1295 | T1475 | T1474 | prep | to,ascribed |
| R1297 | T1476 | T1475 | pobj | them,to |
| R1502 | T1742 | T1744 | nummod | "Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 ",culture |
| R1503 | T1743 | T1744 | compound | Cell,culture |
| R1504 | T1744 | T1744 | ROOT | culture,culture |
| R1505 | T1745 | T1744 | punct | ",",culture |
| R1506 | T1746 | T1747 | compound | transient,transfections |
| R1507 | T1747 | T1744 | appos | transfections,culture |
| R1508 | T1748 | T1747 | cc | and,transfections |
| R1509 | T1749 | T1750 | compound | cell,stimulation |
| R1510 | T1750 | T1747 | conj | stimulation,transfections |
| R1511 | T1751 | T1752 | det | The,generation |
| R1512 | T1752 | T1765 | nsubj | generation,/ |
| R1513 | T1753 | T1752 | punct | ",",generation |
| R1514 | T1754 | T1752 | conj | culture,generation |
| R1515 | T1755 | T1754 | cc | and,culture |
| R1046 | T1160 | T1161 | auxpass | been,proposed |
| R1047 | T1161 | T1161 | ROOT | proposed,proposed |
| R1048 | T1162 | T1163 | aux | to,regulate |
| R1049 | T1163 | T1161 | xcomp | regulate,proposed |
| R1128 | T1242 | T1243 | nummod | 23,] |
| R1129 | T1243 | T1243 | ROOT | ],] |
| R1130 | T1244 | T1223 | punct | .,linked |
| R1516 | T1756 | T1754 | conj | activation,culture |
| R1517 | T1757 | T1752 | prep | of,generation |
| R1518 | T1758 | T1761 | nummod | PKD1,− |
| R175 | T212 | T215 | compound | Protein,enzymes |
| R176 | T213 | T215 | compound | kinase,enzymes |
| R177 | T214 | T215 | compound | D,enzymes |
| R178 | T215 | T216 | nsubj | enzymes,are |
| R179 | T216 | T216 | ROOT | are,are |
| R180 | T217 | T216 | acomp | dispensable,are |
| R181 | T218 | T217 | prep | for,dispensable |
| R182 | T219 | T226 | nmod | proliferation,activity |
| R183 | T220 | T219 | punct | ",",proliferation |
| R184 | T221 | T219 | conj | survival,proliferation |
| R185 | T222 | T221 | cc | and,survival |
| R186 | T223 | T221 | conj | antigen,survival |
| R187 | T224 | T226 | amod | receptor-regulated,activity |
| R188 | T225 | T226 | compound | NFκB,activity |
| R189 | T226 | T218 | pobj | activity,for |
| R190 | T227 | T226 | prep | in,activity |
| R191 | T228 | T229 | amod | vertebrate,B-cells |
| R192 | T229 | T227 | pobj | B-cells,in |
| R193 | T230 | T231 | aux | To,investigate |
| R194 | T231 | T216 | advcl | investigate,are |
| R195 | T232 | T233 | det | the,importance |
| R196 | T233 | T231 | dobj | importance,investigate |
| R197 | T234 | T233 | prep | of,importance |
| R198 | T235 | T234 | pobj | protein,of |
| R199 | T236 | T237 | compound | kinase,D |
| R200 | T237 | T235 | appos | D,protein |
| R201 | T238 | T237 | punct | (,D |
| R202 | T239 | T237 | appos | PKD,D |
| R203 | T240 | T237 | punct | ),D |
UBERON-AE
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T9 | 117-127 | http://purl.obolibrary.org/obo/UBERON_3010224 | denotes | vertebrate |
GO-BP
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T504 | 1364-1374 | http://purl.obolibrary.org/obo/GO_0023052 | denotes | signalling |
| T505 | 2014-2021 | http://purl.obolibrary.org/obo/GO_0023052 | denotes | signals |
| T506 | 1364-1382 | http://purl.obolibrary.org/obo/GO_0007165 | denotes | signalling pathway |
| T507 | 1562-1565 | http://purl.obolibrary.org/obo/GO_0004697 | denotes | PKC |
| T508 | 1941-1949 | http://purl.obolibrary.org/obo/GO_0007349 | denotes | cellular |
| T509 | 3342-3350 | http://purl.obolibrary.org/obo/GO_0007349 | denotes | cellular |
| T510 | 3625-3633 | http://purl.obolibrary.org/obo/GO_0007349 | denotes | cellular |
| T511 | 3681-3689 | http://purl.obolibrary.org/obo/GO_0007349 | denotes | cellular |
| T512 | 1971-1989 | http://purl.obolibrary.org/obo/GO_0008283 | denotes | cell proliferation |
| T513 | 2044-2055 | http://purl.obolibrary.org/obo/GO_0032502 | denotes | development |
| T514 | 2172-2185 | http://purl.obolibrary.org/obo/GO_0007155 | denotes | cell adhesion |
| T515 | 2190-2203 | http://purl.obolibrary.org/obo/GO_0048870 | denotes | cell motility |
| T516 | 2298-2311 | http://purl.obolibrary.org/obo/GO_0006351 | denotes | transcription |
| T517 | 2660-2673 | http://purl.obolibrary.org/obo/GO_0006351 | denotes | transcription |
| T518 | 3489-3504 | http://purl.obolibrary.org/obo/GO_0006351 | denotes | transcriptional |
| T519 | 2451-2469 | http://purl.obolibrary.org/obo/GO_0006986 | denotes | heat shock protein |
| T520 | 2451-2469 | http://purl.obolibrary.org/obo/GO_0034620 | denotes | heat shock protein |
| T521 | 2451-2469 | http://purl.obolibrary.org/obo/GO_0042026 | denotes | heat shock protein |
| T522 | 2493-2507 | http://purl.obolibrary.org/obo/GO_0016477 | denotes | cell migration |
| T523 | 3132-3142 | http://purl.obolibrary.org/obo/GO_0065007 | denotes | regulation |
| T524 | 3217-3232 | http://purl.obolibrary.org/obo/GO_0016310 | denotes | phosphorylation |
| T525 | 3390-3416 | http://purl.obolibrary.org/obo/GO_1902883 | denotes | oxidative stress responses |
| T526 | 3390-3416 | http://purl.obolibrary.org/obo/GO_0006979 | denotes | oxidative stress responses |
| T527 | 3390-3416 | http://purl.obolibrary.org/obo/GO_0097468 | denotes | oxidative stress responses |
| T528 | 3390-3416 | http://purl.obolibrary.org/obo/GO_1902884 | denotes | oxidative stress responses |
| T529 | 3390-3416 | http://purl.obolibrary.org/obo/GO_0034599 | denotes | oxidative stress responses |
| T530 | 3400-3416 | http://purl.obolibrary.org/obo/GO_0006950 | denotes | stress responses |
| T531 | 3625-3643 | http://purl.obolibrary.org/obo/GO_0009987 | denotes | cellular processes |
| T21 | 553-563 | http://purl.obolibrary.org/obo/GO_0065007 | denotes | regulation |
| T22 | 705-720 | http://purl.obolibrary.org/obo/GO_0016310 | denotes | phosphorylation |
| T23 | 839-847 | http://purl.obolibrary.org/obo/GO_0007349 | denotes | cellular |
| T24 | 839-857 | http://purl.obolibrary.org/obo/GO_0009987 | denotes | cellular processes |
| T25 | 912-927 | http://purl.obolibrary.org/obo/GO_0006351 | denotes | transcriptional |
GO-MF
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T1483 | 1562-1565 | http://purl.obolibrary.org/obo/GO_0004697 | denotes | PKC |
| T1484 | 1729-1736 | http://purl.obolibrary.org/obo/GO_0005488 | denotes | binding |
| T1485 | 1755-1762 | http://purl.obolibrary.org/obo/GO_0005488 | denotes | binding |
GO-CC
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T1499 | 2156-2161 | http://purl.obolibrary.org/obo/GO_0005794 | denotes | Golgi |
| T1500 | 3625-3643 | http://purl.obolibrary.org/obo/GO_0042995 | denotes | cellular processes |
| T1816 | 3773-3777 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1817 | 3879-3883 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1486 | 1157-1161 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1487 | 1971-1975 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1488 | 2172-2176 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1489 | 2190-2194 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1490 | 2337-2341 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1491 | 2493-2497 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1492 | 2512-2516 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1493 | 2781-2785 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T1494 | 2930-2934 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T399 | 130-135 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
| T400 | 376-381 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
| T401 | 731-736 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
| T402 | 793-797 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T403 | 957-961 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T404 | 1023-1027 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cell |
| T405 | 839-857 | http://purl.obolibrary.org/obo/GO_0042995 | denotes | cellular processes |
| T1495 | 3148-3153 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
| T1496 | 3238-3243 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
| T1497 | 3270-3275 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
| T1498 | 3422-3427 | http://purl.obolibrary.org/obo/GO_0005623 | denotes | cells |
sentences
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T17 | 615-646 | Sentence | denotes | Mol. Cell Biol. 26, 1569–1577]. |
| T489 | 635-1151 | Sentence | denotes | 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. |
| T490 | 1152-1351 | Sentence | denotes | Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. |
| T491 | 1352-1665 | Sentence | denotes | A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. |
| T492 | 1666-1888 | Sentence | denotes | The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. |
| T493 | 1889-2061 | Sentence | denotes | PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. |
| T494 | 2062-2223 | Sentence | denotes | Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). |
| T495 | 2224-2386 | Sentence | denotes | In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. |
| T496 | 2387-2531 | Sentence | denotes | Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. |
| T497 | 2532-2712 | Sentence | denotes | An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. |
| T498 | 2713-3066 | Sentence | denotes | To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. |
| T499 | 3067-3158 | Sentence | denotes | Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. |
| T500 | 3159-3244 | Sentence | denotes | Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. |
| T501 | 3245-3312 | Sentence | denotes | However, PKD-null DT40 B cells are viable and proliferate normally. |
| T502 | 3313-3514 | Sentence | denotes | Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. |
| T503 | 3515-3698 | Sentence | denotes | Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. |
| T1670 | 0-3789 | Sentence | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell culture, transient transfections and cell stimulation |
| T1671 | 3790-3925 | Sentence | denotes | The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. |
| T1672 | 3926-4038 | Sentence | denotes | Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. |
| T1673 | 4039-4117 | Sentence | denotes | Chloramphenicol acetyl transferase assays have been described previously [29]. |
| T1864 | 0-4137 | Sentence | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells Abstract To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B-cell biology. 1 Introduction The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. However, PKD-null DT40 B cells are viable and proliferate normally. Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. 2 Materials and methods 2.1 Cell culture, transient transfections and cell stimulation The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. Chloramphenicol acetyl transferase assays have been described previously [29]. 2.2 sIgM staining |
| T14 | 0-135 | Sentence | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells |
| T15 | 147-263 | Sentence | denotes | To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. |
| T16 | 264-614 | Sentence | denotes | Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. |
| T18 | 647-737 | Sentence | denotes | We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. |
| T19 | 738-979 | Sentence | denotes | However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. |
| T20 | 980-1036 | Sentence | denotes | Thus, PKDs have a selective role in DT40 B-cell biology. |
| T1 | 0-135 | Sentence | denotes | Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor-regulated NFκB activity in vertebrate B-cells |
| T2 | 138-146 | Sentence | denotes | Abstract |
| T3 | 147-263 | Sentence | denotes | To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD-null DT40 B-lymphocyte cell line. |
| T4 | 264-614 | Sentence | denotes | Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B-cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. |
| T5 | 615-619 | Sentence | denotes | Mol. |
| T6 | 620-630 | Sentence | denotes | Cell Biol. |
| T7 | 631-646 | Sentence | denotes | 26, 1569–1577]. |
| T8 | 647-737 | Sentence | denotes | We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B-cells. |
| T9 | 738-979 | Sentence | denotes | However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. |
| T10 | 980-1036 | Sentence | denotes | Thus, PKDs have a selective role in DT40 B-cell biology. |
| T11 | 1038-1053 | Sentence | denotes | 1 Introduction |
| T12 | 1054-1151 | Sentence | denotes | The protein kinase D (PKD) serine/threonine kinase family has three members: PKD1, PKD2 and PKD3. |
| T13 | 1152-1351 | Sentence | denotes | Most cell types express at least two PKD isoforms but PKD enzymes are especially highly expressed in haematopoietic cells, where they are activated in response to antigen receptors stimulation [2,3]. |
| T14 | 1352-1665 | Sentence | denotes | A conserved signalling pathway linking antigen receptors to PKDs involves the activation of PLCγ and the subsequent production of diacylglycerol (DAG) which stimulates classical and/or novel protein kinase Cs (PKC) that phosphorylate two key regulatory serine residues in the activation loop of PKD kinases [3–6]. |
| T15 | 1666-1888 | Sentence | denotes | The N-terminal regulatory region of PKD enzymes contains a DAG binding domain and direct binding of DAG also contributes to PKD1 activation [7] as well as regulating the spatial location of PKD enzymes within cells [8–12]. |
| T16 | 1889-2061 | Sentence | denotes | PKD enzymes have been proposed to regulate numerous cellular functions, including cell proliferation [13–16], anti-apoptotic signals [17,18] and thymocyte development [19]. |
| T17 | 2062-2223 | Sentence | denotes | Expression of mutant catalytically inactive and constitutively activated PKDs can also modify Golgi function, cell adhesion and cell motility (reviewed in [20]). |
| T18 | 2224-2386 | Sentence | denotes | In particular, PKDs have been widely linked to the activation of the NFκB transcription factor and in regulating cell survival during oxidative stress [17,21–23]. |
| T19 | 2387-2531 | Sentence | denotes | Another recently proposed PKD1 substrate is HSP27 [24], a small heat shock protein involved in regulating cell migration and cell survival [25]. |
| T20 | 2532-2712 | Sentence | denotes | An essential role for PKD enzymes in regulating class II histone deacetylases (HDACs), enzymes that repress MEF2-dependent gene transcription, has also been demonstrated [1,26–28]. |
| T21 | 2713-3066 | Sentence | denotes | To investigate the biological role of PKDs we have generated DT40 B cell lines that lack expression of one or more members of the PKD family [1], allowing us to investigate the function(s) of PKD isoforms following B cell antigen receptor (BCR) stimulation, as well addressing the issue of functional redundancy between the different PKD family members. |
| T22 | 3067-3158 | Sentence | denotes | Previous studies have shown that PKDs are indispensable for HDAC regulation in B cells [1]. |
| T23 | 3159-3244 | Sentence | denotes | Herein we show that PKDs are also indispensable for HSP27 phosphorylation in B cells. |
| T24 | 3245-3312 | Sentence | denotes | However, PKD-null DT40 B cells are viable and proliferate normally. |
| T25 | 3313-3514 | Sentence | denotes | Moreover, loss of the entire cellular pool of PKD does not critically affect oxidative stress responses in B cells nor do PKD kinases play an essential role in regulating NFκB transcriptional activity. |
| T26 | 3515-3698 | Sentence | denotes | Together, these findings reveal that in B lymphocytes, PKD kinases are not critical regulators of many of the cellular processes previously ascribed to them in other cellular systems. |
| T27 | 3700-3724 | Sentence | denotes | 2 Materials and methods |
| T28 | 3726-3789 | Sentence | denotes | 2.1 Cell culture, transient transfections and cell stimulation |
| T29 | 3790-3925 | Sentence | denotes | The generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. |
| T30 | 3926-4038 | Sentence | denotes | Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. |
| T31 | 4039-4117 | Sentence | denotes | Chloramphenicol acetyl transferase assays have been described previously [29]. |
| T32 | 4119-4137 | Sentence | denotes | 2.2 sIgM staining |
ICD10
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T1482 | 2456-2461 | http://purl.bioontology.org/ontology/ICD10/R57.9 | denotes | shock |
events-check-again
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T425 | 678-686 | Positive_regulation | denotes | required |
| T426 | 690-698 | Regulation | denotes | regulate |
| T427 | 699-704 | Protein | denotes | HSP27 |
| T428 | 705-720 | Phosphorylation | denotes | phosphorylation |
| T1575 | 1131-1135 | Protein | denotes | PKD1 |
| T1576 | 1137-1141 | Protein | denotes | PKD2 |
| T1577 | 1146-1150 | Protein | denotes | PKD3 |
| T1578 | 1755-1762 | Binding | denotes | binding |
| T1579 | 1775-1786 | Positive_regulation | denotes | contributes |
| T1580 | 1790-1794 | Protein | denotes | PKD1 |
| T1581 | 1795-1805 | Positive_regulation | denotes | activation |
| T1582 | 2413-2417 | Protein | denotes | PKD1 |
| T1583 | 2431-2436 | Protein | denotes | HSP27 |
| T1584 | 2640-2644 | Protein | denotes | MEF2 |
| T1585 | 3193-3206 | Positive_regulation | denotes | indispensable |
| T1586 | 3211-3216 | Protein | denotes | HSP27 |
| T1587 | 3217-3232 | Phosphorylation | denotes | phosphorylation |
| T1851 | 3832-3836 | Protein | denotes | PKD1 |
| T1852 | 3841-3845 | Protein | denotes | PKD3 |
| T1853 | 3853-3857 | Protein | denotes | PKD1 |
| T1854 | 3858-3859 | Protein | denotes | 3 |
| T1855 | 3863-3871 | Negative_regulation | denotes | knockout |
| T1856 | 3863-3871 | Negative_regulation | denotes | knockout |
| T1857 | 3863-3871 | Negative_regulation | denotes | knockout |
| T1858 | 3863-3871 | Negative_regulation | denotes | knockout |
| R410 | T426 | T425 | themeOf | regulate,required |
| R411 | T427 | T428 | themeOf | HSP27,phosphorylation |
| R412 | T428 | T426 | themeOf | phosphorylation,regulate |
| R1400 | T1578 | T1579 | causeOf | binding,contributes |
| R1402 | T1580 | T1578 | themeOf | PKD1,binding |
| R1403 | T1580 | T1581 | themeOf | PKD1,activation |
| R1404 | T1581 | T1579 | themeOf | activation,contributes |
| R1405 | T1586 | T1587 | themeOf | HSP27,phosphorylation |
| R1406 | T1587 | T1585 | themeOf | phosphorylation,indispensable |
| R1592 | T1851 | T1855 | themeOf | PKD1,knockout |
| R1593 | T1852 | T1856 | themeOf | PKD3,knockout |
| R1594 | T1853 | T1857 | themeOf | PKD1,knockout |
| R1595 | T1854 | T1858 | themeOf | 3,knockout |
bionlp-st-ge-2016-reference-tees
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T429 | 0-24 | Protein | denotes | Protein kinase D enzymes |
| T430 | 100-104 | Protein | denotes | NFκB |
| T431 | 81-99 | Regulation | denotes | receptor-regulated |
| T432 | 180-196 | Protein | denotes | protein kinase D |
| T433 | 198-201 | Protein | denotes | PKD |
| T434 | 226-229 | Protein | denotes | PKD |
| T435 | 294-298 | Protein | denotes | PKDs |
| T436 | 336-365 | Protein | denotes | class II histone deacetylases |
| T437 | 514-545 | Protein | denotes | protein kinase D family kinases |
| T438 | 567-596 | Protein | denotes | class II histone deacetylases |
| T439 | 553-563 | Regulation | denotes | regulation |
| T440 | 664-668 | Protein | denotes | PKDs |
| T441 | 699-704 | Protein | denotes | HSP27 |
| T442 | 705-720 | Phosphorylation | denotes | phosphorylation |
| T443 | 690-698 | Regulation | denotes | regulate |
| T444 | 678-686 | Positive_regulation | denotes | required |
| T445 | 805-816 | Protein | denotes | PKD enzymes |
| T446 | 907-911 | Protein | denotes | NFκB |
| T447 | 955-978 | Protein | denotes | B cell antigen receptor |
| T448 | 824-832 | Regulation | denotes | regulate |
| T449 | 986-990 | Protein | denotes | PKDs |
| T1588 | 1058-1074 | Protein | denotes | protein kinase D |
| T1589 | 1076-1079 | Protein | denotes | PKD |
| T1590 | 1081-1111 | Protein | denotes | serine/threonine kinase family |
| T1591 | 1131-1135 | Protein | denotes | PKD1 |
| T1592 | 1137-1141 | Protein | denotes | PKD2 |
| T1593 | 1146-1150 | Protein | denotes | PKD3 |
| T1594 | 1189-1192 | Protein | denotes | PKD |
| T1595 | 1206-1209 | Protein | denotes | PKD |
| T1596 | 1168-1175 | Gene_expression | denotes | express |
| T1597 | 1240-1249 | Gene_expression | denotes | expressed |
| T1598 | 1412-1416 | Protein | denotes | PKDs |
| T1599 | 1444-1448 | Protein | denotes | PLCγ |
| T1600 | 1543-1560 | Protein | denotes | protein kinase Cs |
| T1601 | 1562-1565 | Protein | denotes | PKC |
| T1602 | 1647-1658 | Protein | denotes | PKD kinases |
| T1603 | 1509-1519 | Positive_regulation | denotes | stimulates |
| T1604 | 1509-1519 | Positive_regulation | denotes | stimulates |
| T1605 | 1572-1585 | Phosphorylation | denotes | phosphorylate |
| T1606 | 1572-1585 | Phosphorylation | denotes | phosphorylate |
| T1607 | 1702-1713 | Protein | denotes | PKD enzymes |
| T1608 | 1766-1769 | Protein | denotes | DAG |
| T1609 | 1790-1794 | Protein | denotes | PKD1 |
| T1610 | 1856-1867 | Protein | denotes | PKD enzymes |
| T1611 | 1755-1762 | Binding | denotes | binding |
| T1612 | 1795-1805 | Positive_regulation | denotes | activation |
| T1613 | 1844-1852 | Localization | denotes | location |
| T1614 | 1775-1786 | Positive_regulation | denotes | contributes |
| T1615 | 1821-1831 | Regulation | denotes | regulating |
| T1616 | 1889-1892 | Protein | denotes | PKD |
| T1617 | 2135-2139 | Protein | denotes | PKDs |
| T1618 | 2062-2072 | Gene_expression | denotes | Expression |
| T1619 | 2125-2134 | Positive_regulation | denotes | activated |
| T1620 | 2125-2134 | Positive_regulation | denotes | activated |
| T1621 | 2239-2243 | Protein | denotes | PKDs |
| T1622 | 2293-2318 | Protein | denotes | NFκB transcription factor |
| T1623 | 2275-2285 | Positive_regulation | denotes | activation |
| T1624 | 2413-2417 | Protein | denotes | PKD1 |
| T1625 | 2554-2565 | Protein | denotes | PKD enzymes |
| T1626 | 2580-2609 | Protein | denotes | class II histone deacetylases |
| T1627 | 2611-2616 | Protein | denotes | HDACs |
| T1628 | 2640-2644 | Protein | denotes | MEF2 |
| T1629 | 2569-2579 | Regulation | denotes | regulating |
| T1630 | 2569-2579 | Regulation | denotes | regulating |
| T1631 | 2751-2755 | Protein | denotes | PKDs |
| T1632 | 2843-2846 | Protein | denotes | PKD |
| T1633 | 2905-2908 | Protein | denotes | PKD |
| T1634 | 2928-2951 | Protein | denotes | B cell antigen receptor |
| T1635 | 2953-2956 | Protein | denotes | BCR |
| T1636 | 3047-3065 | Protein | denotes | PKD family members |
| T1637 | 3100-3104 | Protein | denotes | PKDs |
| T1638 | 3127-3131 | Protein | denotes | HDAC |
| T1639 | 3132-3142 | Regulation | denotes | regulation |
| T1640 | 3179-3183 | Protein | denotes | PKDs |
| T1641 | 3211-3216 | Protein | denotes | HSP27 |
| T1642 | 3217-3232 | Phosphorylation | denotes | phosphorylation |
| T1643 | 3193-3206 | Positive_regulation | denotes | indispensable |
| T1644 | 3254-3257 | Protein | denotes | PKD |
| T1645 | 3359-3362 | Protein | denotes | PKD |
| T1646 | 3435-3446 | Protein | denotes | PKD kinases |
| T1647 | 3484-3488 | Protein | denotes | NFκB |
| T1648 | 3323-3327 | Negative_regulation | denotes | loss |
| T1649 | 3570-3581 | Protein | denotes | PKD kinases |
| T1859 | 3832-3836 | Protein | denotes | PKD1 |
| T1860 | 3841-3845 | Protein | denotes | PKD3 |
| T1861 | 3853-3857 | Protein | denotes | PKD1 |
| T1862 | 4039-4073 | Protein | denotes | Chloramphenicol acetyl transferase |
| R1413 | T1609 | T1612 | themeOf | PKD1,activation |
| R1414 | T1610 | T1613 | themeOf | PKD enzymes,location |
| R1415 | T1611 | T1614 | causeOf | binding,contributes |
| R1416 | T1612 | T1614 | themeOf | activation,contributes |
| R1417 | T1613 | T1615 | themeOf | location,regulating |
| R1418 | T1617 | T1618 | themeOf | PKDs,Expression |
| R1419 | T1617 | T1619 | themeOf | PKDs,activated |
| R1420 | T1618 | T1620 | themeOf | Expression,activated |
| R1421 | T1622 | T1623 | themeOf | NFκB transcription factor,activation |
| R1422 | T1626 | T1629 | themeOf | class II histone deacetylases,regulating |
| R1423 | T1627 | T1630 | themeOf | HDACs,regulating |
| R1424 | T1638 | T1639 | themeOf | HDAC,regulation |
| R1425 | T1640 | T1643 | causeOf | PKDs,indispensable |
| R1426 | T1641 | T1642 | themeOf | HSP27,phosphorylation |
| R1427 | T1642 | T1643 | themeOf | phosphorylation,indispensable |
| R1428 | T1645 | T1648 | themeOf | PKD,loss |
| R1401 | T1594 | T1596 | themeOf | PKD,express |
| R1407 | T1595 | T1597 | themeOf | PKD,expressed |
| R1408 | T1600 | T1603 | themeOf | protein kinase Cs,stimulates |
| R1409 | T1600 | T1605 | themeOf | protein kinase Cs,phosphorylate |
| R1410 | T1601 | T1604 | themeOf | PKC,stimulates |
| R1411 | T1601 | T1606 | themeOf | PKC,phosphorylate |
| R1412 | T1608 | T1611 | themeOf | DAG,binding |
| R362 | T430 | T431 | themeOf | NFκB,receptor-regulated |
| R363 | T438 | T439 | themeOf | class II histone deacetylases,regulation |
| R364 | T440 | T443 | causeOf | PKDs,regulate |
| R365 | T440 | T444 | causeOf | PKDs,required |
| R366 | T441 | T442 | themeOf | HSP27,phosphorylation |
| R367 | T442 | T443 | themeOf | phosphorylation,regulate |
| R368 | T443 | T444 | themeOf | regulate,required |
| R369 | T445 | T448 | causeOf | PKD enzymes,regulate |
| R370 | T447 | T448 | themeOf | B cell antigen receptor,regulate |
bionlp-st-ge-2016-reference
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T10 | 678-686 | Positive_regulation | denotes | required |
| T11 | 690-698 | Regulation | denotes | regulate |
| T12 | 699-704 | Protein | denotes | HSP27 |
| T13 | 705-720 | Phosphorylation | denotes | phosphorylation |
| T476 | 1131-1135 | Protein | denotes | PKD1 |
| T477 | 1137-1141 | Protein | denotes | PKD2 |
| T478 | 1146-1150 | Protein | denotes | PKD3 |
| T479 | 1755-1762 | Binding | denotes | binding |
| T480 | 1775-1786 | Positive_regulation | denotes | contributes |
| T481 | 1790-1794 | Protein | denotes | PKD1 |
| T482 | 1795-1805 | Positive_regulation | denotes | activation |
| T483 | 2413-2417 | Protein | denotes | PKD1 |
| T484 | 2431-2436 | Protein | denotes | HSP27 |
| T485 | 2640-2644 | Protein | denotes | MEF2 |
| T486 | 3193-3206 | Positive_regulation | denotes | indispensable |
| T487 | 3211-3216 | Protein | denotes | HSP27 |
| T488 | 3217-3232 | Phosphorylation | denotes | phosphorylation |
| T1662 | 3832-3836 | Protein | denotes | PKD1 |
| T1663 | 3841-3845 | Protein | denotes | PKD3 |
| T1664 | 3853-3857 | Protein | denotes | PKD1 |
| T1665 | 3858-3859 | Protein | denotes | 3 |
| T1666 | 3863-3871 | Negative_regulation | denotes | knockout |
| T1667 | 3863-3871 | Negative_regulation | denotes | knockout |
| T1668 | 3863-3871 | Negative_regulation | denotes | knockout |
| T1669 | 3863-3871 | Negative_regulation | denotes | knockout |
| R371 | T11 | T10 | themeOf | regulate,required |
| R372 | T12 | T13 | themeOf | HSP27,phosphorylation |
| R373 | T13 | T11 | themeOf | phosphorylation,regulate |
| R424 | T479 | T480 | causeOf | binding,contributes |
| R425 | T481 | T479 | themeOf | PKD1,binding |
| R426 | T481 | T482 | themeOf | PKD1,activation |
| R427 | T482 | T480 | themeOf | activation,contributes |
| R428 | T487 | T488 | themeOf | HSP27,phosphorylation |
| R429 | T488 | T486 | themeOf | phosphorylation,indispensable |
| R1433 | T1662 | T1666 | themeOf | PKD1,knockout |
| R1434 | T1663 | T1667 | themeOf | PKD3,knockout |
| R1435 | T1664 | T1668 | themeOf | PKD1,knockout |
| R1436 | T1665 | T1669 | themeOf | 3,knockout |
bionlp-st-ge-2016-uniprot
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T1737 | 3853-3857 | P98161 | denotes | PKD1 |
| T986 | 1131-1135 | P98161 | denotes | PKD1 |
| T987 | 1146-1150 | Q99853 | denotes | PKD3 |
| T988 | 1790-1794 | P98161 | denotes | PKD1 |
| T989 | 2413-2417 | P98161 | denotes | PKD1 |
| T990 | 2431-2436 | P04792 | denotes | HSP27 |
| T991 | 2953-2956 | P11274 | denotes | BCR |
| T992 | 3211-3216 | P04792 | denotes | HSP27 |
| T1735 | 3832-3836 | P98161 | denotes | PKD1 |
| T1736 | 3841-3845 | Q99853 | denotes | PKD3 |
| T204 | 699-704 | P04792 | denotes | HSP27 |
test2
| Id | Subject | Object | Predicate | Lexical cue |
|---|---|---|---|---|
| T5 | 678-686 | Positive_regulation | denotes | required |
| T6 | 690-698 | Regulation | denotes | regulate |
| T7 | 699-704 | Protein | denotes | HSP27 |
| T8 | 705-720 | Phosphorylation | denotes | phosphorylation |
| T463 | 1131-1135 | Protein | denotes | PKD1 |
| T464 | 1137-1141 | Protein | denotes | PKD2 |
| T465 | 1146-1150 | Protein | denotes | PKD3 |
| T466 | 1755-1762 | Binding | denotes | binding |
| T467 | 1775-1786 | Positive_regulation | denotes | contributes |
| T468 | 1790-1794 | Protein | denotes | PKD1 |
| T469 | 1795-1805 | Positive_regulation | denotes | activation |
| T470 | 2413-2417 | Protein | denotes | PKD1 |
| T471 | 2431-2436 | Protein | denotes | HSP27 |
| T472 | 2640-2644 | Protein | denotes | MEF2 |
| T473 | 3193-3206 | Positive_regulation | denotes | indispensable |
| T474 | 3211-3216 | Protein | denotes | HSP27 |
| T475 | 3217-3232 | Phosphorylation | denotes | phosphorylation |
| T1658 | 3832-3836 | Protein | denotes | PKD1 |
| T1659 | 3841-3845 | Protein | denotes | PKD3 |
| T1660 | 3853-3857 | Protein | denotes | PKD1 |
| T1661 | 3858-3859 | Protein | denotes | 3 |
| R4 | T6 | T5 | themeOf | regulate,required |
| R5 | T7 | T8 | themeOf | HSP27,phosphorylation |
| R6 | T8 | T6 | themeOf | phosphorylation,regulate |
| R419 | T466 | T467 | causeOf | binding,contributes |
| R420 | T468 | T469 | themeOf | PKD1,activation |
| R421 | T469 | T467 | themeOf | activation,contributes |
| R422 | T474 | T475 | themeOf | HSP27,phosphorylation |
| R423 | T475 | T473 | themeOf | phosphorylation,indispensable |