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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
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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
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T2288 0-4551 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 staining DT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice. The cells were washed twice and fluorescent intensity was analysed by flow cytometry. All results shown are representative of at two to four independent experiments unless otherwise indicated. 3 Results 3.1 Loss
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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
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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"
R1598 T1872 T1870 arg1Of cells,DT40
R1599 T1872 T1871 arg1Of cells,B
R1600 T1872 T1873 arg1Of cells,(
R1601 T1872 T1880 arg2Of cells,resuspended
R1604 T1875 T1873 arg2Of cells,(
R1606 T1878 T1873 arg3Of ),(
R1607 T1880 T1879 arg2Of resuspended,were
R1608 T1880 T1881 arg1Of resuspended,in
R1609 T1883 T1881 arg2Of buffer,in
R1610 T1883 T1882 arg1Of buffer,200 μl
R1611 T1883 T1884 arg1Of buffer,(
R1613 T1887 T1884 arg2Of media,(
R1614 T1887 T1885 arg1Of media,RPMI
R1615 T1887 T1886 arg1Of media,1640
R1616 T1887 T1888 arg1Of media,","
R1618 T1893 T1888 arg2Of serum,","
R1619 T1893 T1889 arg1Of serum,1
R1622 T1894 T1884 arg3Of ),(
R1653 T1924 T1923 arg2Of representative,are
R1654 T1924 T1926 arg1Of representative,at
R1655 T1926 T1925 arg1Of at,of
R1656 T1927 T1928 arg1Of two,to
R1657 T1929 T1928 arg2Of four,to
R1658 T1931 T1926 arg2Of experiments,at
R1659 T1931 T1927 arg1Of experiments,two
R1660 T1931 T1930 arg1Of experiments,independent
R1661 T1934 T1932 arg2Of indicated,unless
R1662 T1934 T1933 arg1Of indicated,otherwise
R1597 T1872 T1879 arg1Of cells,were
R1602 T1875 T1874 arg1Of cells,2 × 106
R1603 T1875 T1876 arg1Of cells,per
R1605 T1877 T1876 arg2Of point,per
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
R1612 T1883 T1895 arg1Of buffer,containing
R1617 T1889 T1890 arg1Of 1,%
R1620 T1893 T1891 arg1Of serum,foetal
R1621 T1893 T1892 arg1Of serum,calf
R1623 T1899 T1895 arg2Of antibody,containing
R1624 T1899 T1896 arg1Of antibody,anti-chicken
R1625 T1899 T1897 arg1Of antibody,M1
R1626 T1899 T1898 arg1Of antibody,monoclonal
R1627 T1899 T1900 arg2Of antibody,conjugated
R1628 T1900 T1901 arg1Of conjugated,to
R1629 T1902 T1901 arg2Of FITC,to
R1630 T1902 T1903 arg1Of FITC,for
R1631 T1904 T1903 arg2Of 20 min,for
R1632 T1904 T1905 arg1Of 20 min,on
R1633 T1906 T1905 arg2Of ice,on
R1634 T1908 T1907 arg1Of cells,The
R1635 T1908 T1909 arg1Of cells,were
R1636 T1908 T1910 arg2Of cells,washed
R1637 T1910 T1909 arg2Of washed,were
R1638 T1910 T1911 arg1Of washed,twice
R1639 T1910 T1912 arg1Of washed,and
R1640 T1914 T1913 arg1Of intensity,fluorescent
R1641 T1914 T1915 arg1Of intensity,was
R1642 T1914 T1916 arg2Of intensity,analysed
R1643 T1916 T1912 arg2Of analysed,and
R1644 T1916 T1915 arg2Of analysed,was
R1645 T1919 T1916 arg1Of cytometry,analysed
R1646 T1919 T1917 arg2Of cytometry,by
R1647 T1919 T1918 arg1Of cytometry,flow
R1648 T1921 T1920 arg1Of results,All
R1649 T1921 T1922 arg2Of results,shown
R1650 T1921 T1923 arg1Of results,are
R1651 T1921 T1924 arg1Of results,representative
R1652 T1923 T1932 arg1Of are,unless

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
T1942 4138-4142 NNP denotes DT40
T1943 4143-4144 NNP denotes B
T1944 4145-4150 NNS denotes cells
T1945 4151-4152 -LRB- denotes (
T1946 4152-4153 CD denotes 2
T1947 4154-4155 CD denotes ×
T1948 4156-4159 CD denotes 106
T1949 4160-4165 NNS denotes cells
T1950 4166-4169 IN denotes per
T1951 4170-4175 NN denotes point
T1952 4175-4176 -RRB- denotes )
T1953 4177-4181 VBD denotes were
T1954 4182-4193 VBN denotes resuspended
T1955 4194-4196 IN denotes in
T1956 4197-4200 CD denotes 200
T1957 4201-4203 NN denotes μl
T1958 4204-4210 NN denotes buffer
T1959 4211-4212 -LRB- denotes (
T1960 4212-4216 NNP denotes RPMI
T1961 4217-4221 CD denotes 1640
T1962 4222-4227 NNS denotes media
T1963 4227-4228 , denotes ,
T1964 4229-4230 CD denotes 1
T1965 4230-4231 NN denotes %
T1966 4232-4238 JJ denotes foetal
T1967 4239-4243 NN denotes calf
T1968 4244-4249 NN denotes serum
T1969 4249-4250 -RRB- denotes )
T1970 4251-4261 VBG denotes containing
T1971 4262-4274 JJ denotes anti-chicken
T1972 4275-4277 CD denotes M1
T1973 4278-4288 JJ denotes monoclonal
T1974 4289-4297 NN denotes antibody
T1975 4298-4308 VBN denotes conjugated
T1976 4309-4311 TO denotes to
T1977 4312-4316 NNP denotes FITC
T1978 4317-4320 IN denotes for
T1979 4321-4323 CD denotes 20
T1980 4324-4327 NN denotes min
T1981 4328-4330 IN denotes on
T1982 4331-4334 NN denotes ice
T1983 4334-4335 . denotes .
T1984 4336-4339 DT denotes The
T1985 4340-4345 NNS denotes cells
T1986 4346-4350 VBD denotes were
T1987 4351-4357 VBN denotes washed
T1988 4358-4363 RB denotes twice
T1989 4364-4367 CC denotes and
T1990 4368-4379 JJ denotes fluorescent
T1991 4380-4389 NN denotes intensity
T1992 4390-4393 VBD denotes was
T1993 4394-4402 VBN denotes analysed
T1994 4403-4405 IN denotes by
T1995 4406-4410 NN denotes flow
T1996 4411-4420 NN denotes cytometry
T1997 4420-4421 . denotes .
T1998 4422-4425 DT denotes All
T1999 4426-4433 NNS denotes results
T2000 4434-4439 VBN denotes shown
T2001 4440-4443 VBP denotes are
T2002 4444-4458 NN denotes representative
T2003 4459-4461 IN denotes of
T2004 4462-4464 IN denotes at
T2005 4465-4468 CD denotes two
T2006 4469-4471 TO denotes to
T2007 4472-4476 CD denotes four
T2008 4477-4488 JJ denotes independent
T2009 4489-4500 NNS denotes experiments
T2010 4501-4507 IN denotes unless
T2011 4508-4517 RB denotes otherwise
T2012 4518-4527 VBN denotes indicated
T2013 4527-4528 . denotes .
T2730 0-4547 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 sIgM staining DT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice. The cells were washed twice and fluorescent intensity was analysed by flow cytometry. All results shown are representative of at two to four independent experiments unless otherwise indicated. 3 Results 3.1
T2731 4547-4551 NN denotes Loss
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
R1664 T1940 T1944 amod sIgM,cells
R1665 T1941 T1944 amod staining,cells
R1666 T1942 T1943 compound DT40,B
R1667 T1943 T1944 compound B,cells
R1668 T1944 T1954 nsubjpass cells,resuspended
R1669 T1945 T1944 punct (,cells
R1670 T1946 T1949 nummod 2,cells
R1671 T1947 T1949 nummod ×,cells
R1677 T1953 T1954 auxpass were,resuspended
R1678 T1954 T1954 ROOT resuspended,resuspended
R1679 T1955 T1954 prep in,resuspended
R1680 T1956 T1958 nummod 200,buffer
R1681 T1957 T1958 compound μl,buffer
R1682 T1958 T1955 pobj buffer,in
R1683 T1959 T1958 punct (,buffer
R1684 T1960 T1962 compound RPMI,media
R1685 T1961 T1962 compound 1640,media
R1686 T1962 T1958 appos media,buffer
R1687 T1963 T1962 punct ",",media
R1688 T1964 T1965 nummod 1,%
R1689 T1965 T1968 nmod %,serum
R1690 T1966 T1968 amod foetal,serum
R1691 T1967 T1968 compound calf,serum
R1692 T1968 T1962 appos serum,media
R1693 T1969 T1962 punct ),media
R1694 T1970 T1958 acl containing,buffer
R1695 T1971 T1975 amod anti-chicken,conjugated
R1696 T1972 T1974 nummod M1,antibody
R1697 T1973 T1974 amod monoclonal,antibody
R1698 T1974 T1975 compound antibody,conjugated
R1699 T1975 T1954 conj conjugated,resuspended
R1700 T1976 T1975 prep to,conjugated
R1701 T1977 T1976 pobj FITC,to
R1702 T1978 T1975 prep for,conjugated
R1703 T1979 T1980 nummod 20,min
R1704 T1980 T1978 pobj min,for
R1705 T1981 T1980 prep on,min
R1706 T1982 T1981 pobj ice,on
R1707 T1983 T1954 punct .,resuspended
R1708 T1984 T1985 det The,cells
R1709 T1985 T1987 nsubjpass cells,washed
R1710 T1986 T1987 auxpass were,washed
R1711 T1987 T1987 ROOT washed,washed
R1712 T1988 T1987 advmod twice,washed
R1713 T1989 T1988 cc and,twice
R1714 T1990 T1991 amod fluorescent,intensity
R1715 T1991 T1993 nsubjpass intensity,analysed
R1716 T1992 T1993 auxpass was,analysed
R1717 T1993 T1987 conj analysed,washed
R1718 T1994 T1993 agent by,analysed
R1719 T1995 T1996 compound flow,cytometry
R1720 T1996 T1994 pobj cytometry,by
R1721 T1997 T1993 punct .,analysed
R1722 T1998 T1999 det All,results
R1723 T1999 T2001 nsubj results,are
R1724 T2000 T1999 acl shown,results
R1725 T2001 T2001 ROOT are,are
R1726 T2002 T2001 attr representative,are
R1727 T2003 T2002 prep of,representative
R1728 T2004 T2001 prep at,are
R1729 T2005 T2007 quantmod two,four
R1730 T2006 T2007 quantmod to,four
R1731 T2007 T2009 nummod four,experiments
R1732 T2008 T2009 amod independent,experiments
R1733 T2009 T2004 pobj experiments,at
R1734 T2010 T2012 mark unless,indicated
R1735 T2011 T2012 advmod otherwise,indicated
R1736 T2012 T2001 advcl indicated,are
R1737 T2013 T2001 punct .,are
R2279 T2730 T2731 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 staining DT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice. The cells were washed twice and fluorescent intensity was analysed by flow cytometry. All results shown are representative of at two to four independent experiments unless otherwise indicated. 3 Results 3.1 ",Loss
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,−
R1672 T1948 T1949 nummod 106,cells
R1673 T1949 T1944 appos cells,cells
R1674 T1950 T1949 prep per,cells
R1675 T1951 T1950 pobj point,per
R1676 T1952 T1944 punct ),cells
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
T1863 4244-4249 http://purl.obolibrary.org/obo/UBERON_0001977 denotes serum

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
T2014 4289-4297 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody

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
T2015 4145-4150 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T2016 4160-4165 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T2017 4340-4345 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T2018 4289-4297 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T2019 4289-4297 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
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
T1865 4138-4335 Sentence denotes DT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice.
T1866 4336-4421 Sentence denotes The cells were washed twice and fluorescent intensity was analysed by flow cytometry.
T1867 4422-4528 Sentence denotes All results shown are representative of at two to four independent experiments unless otherwise indicated.
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
T33 4138-4335 Sentence denotes DT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice.
T34 4336-4421 Sentence denotes The cells were washed twice and fluorescent intensity was analysed by flow cytometry.
T35 4422-4528 Sentence denotes All results shown are representative of at two to four independent experiments unless otherwise indicated.
T36 4530-4540 Sentence denotes 3 Results

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 Speculation
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
T3540 4547-4551 Negative_regulation denotes Loss true
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
T3627 0-4551 Negative_regulation 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 DT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice. The cells were washed twice and fluorescent intensity was analysed by flow cytometry. All results shown are representative of at two to four independent experiments unless otherwise indicated. 3 Results 3.1 Loss
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
T2020 4262-4297 Protein denotes anti-chicken M1 monoclonal antibody
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 Speculation
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
T2177 4547-4551 Negative_regulation denotes Loss true
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