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Id Subject Object Predicate Lexical cue
T26 0-7 NN denotes Protein
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
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
T27 8-14 NN denotes kinase
T28 15-16 NN denotes D
T29 17-24 NN denotes enzymes
T30 25-28 VB denotes are
T31 29-40 JJ denotes dispensable
T32 41-44 IN denotes for
T33 45-58 NN denotes proliferation
T34 58-59 -COMMA- denotes ,
T35 60-68 NN denotes survival
T36 69-72 CC denotes and
T37 73-80 NN denotes antigen
T38 81-99 JJ denotes receptor-regulated
T39 100-104 NN denotes NFκB
T40 105-113 NN denotes activity
T41 114-116 IN denotes in
T42 117-127 NN denotes vertebrate
T43 128-135 NN denotes B-cells
T44 147-149 TO denotes To
T45 150-161 VB denotes investigate
T46 162-165 DT denotes the
T47 166-176 NN denotes importance
T48 177-179 IN denotes of
T49 180-187 NN denotes protein
T50 188-194 NN denotes kinase
T51 195-196 NN denotes D
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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
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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
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T129 584-596 NN denotes deacetylases
T130 597-599 IN denotes in
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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
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
T139 644-645 -RRB- denotes ]
T140 647-649 PRP denotes We
T141 650-653 RB denotes now
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T145 669-672 VB denotes are
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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
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T163 787-792 JJ denotes other
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T165 798-803 NN denotes types
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T168 809-816 NN denotes enzymes
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T172 833-838 JJ denotes basic
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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
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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
T533 1058-1065 NN denotes protein
T534 1066-1072 NN denotes kinase
T535 1073-1074 NN denotes D
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T537 1076-1079 NN denotes PKD
T538 1079-1080 -RRB- denotes )
T539 1081-1097 NN denotes serine/threonine
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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
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T553 1162-1167 NN denotes types
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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
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T579 1323-1332 NN denotes receptors
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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
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T602 1479-1481 IN denotes of
T603 1482-1496 NN denotes diacylglycerol
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T605 1498-1501 NN denotes DAG
T606 1501-1502 -RRB- denotes )
T607 1503-1508 WDT denotes which
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T609 1520-1529 JJ denotes classical
T610 1530-1536 CC denotes and/or
T611 1537-1542 JJ denotes novel
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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
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T644 1725-1728 NN denotes DAG
T645 1729-1736 NN denotes binding
T646 1737-1743 NN denotes domain
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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
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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
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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 ]
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T734 2239-2243 NN denotes PKDs
T735 2244-2248 VB denotes have
T736 2249-2253 VB denotes been
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T743 2289-2292 DT denotes the
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T748 2323-2325 IN denotes in
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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
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T763 2428-2430 VB denotes is
T764 2431-2436 NN denotes HSP27
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T766 2438-2440 CD denotes 24
T767 2440-2441 -RRB- denotes ]
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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
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T789 2554-2557 NN denotes PKD
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T795 2589-2596 NN denotes histone
T796 2597-2609 NN denotes deacetylases
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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
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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
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T826 2779-2780 NN denotes B
T827 2781-2785 NN denotes cell
T828 2786-2791 NN denotes lines
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T830 2797-2801 VB denotes lack
T831 2802-2812 NN denotes expression
T832 2813-2815 IN denotes of
T833 2816-2819 CD denotes one
T834 2820-2822 CC denotes or
T835 2823-2827 JJ denotes more
T836 2828-2835 NN denotes members
T837 2836-2838 IN denotes of
T838 2839-2842 DT denotes the
T839 2843-2846 NN denotes PKD
T840 2847-2853 NN denotes family
T841 2854-2855 -LRB- denotes [
T842 2855-2856 CD denotes 1
T843 2856-2857 -RRB- denotes ]
T844 2857-2858 -COMMA- denotes ,
T845 2859-2867 VB denotes allowing
T846 2868-2870 PRP denotes us
T847 2871-2873 TO denotes to
T848 2874-2885 VB denotes investigate
T849 2886-2889 DT denotes the
T850 2890-2898 NN denotes function
T851 2898-2899 -LRB- denotes (
T852 2899-2900 NN denotes s
T853 2900-2901 -RRB- denotes )
T854 2902-2904 IN denotes of
T855 2905-2908 NN denotes PKD
T856 2909-2917 NN denotes isoforms
T857 2918-2927 VB denotes following
T858 2928-2929 NN denotes B
T859 2930-2934 NN denotes cell
T860 2935-2942 NN denotes antigen
T861 2943-2951 NN denotes receptor
T862 2952-2953 -LRB- denotes (
T863 2953-2956 NN denotes BCR
T864 2956-2957 -RRB- denotes )
T865 2958-2969 NN denotes stimulation
T866 2969-2970 -COMMA- denotes ,
T867 2971-2973 RB denotes as
T868 2974-2978 RB denotes well
T869 2979-2989 VB denotes addressing
T870 2990-2993 DT denotes the
T871 2994-2999 NN denotes issue
T872 3000-3002 IN denotes of
T873 3003-3013 JJ denotes functional
T874 3014-3024 NN denotes redundancy
T875 3025-3032 IN denotes between
T876 3033-3036 DT denotes the
T877 3037-3046 JJ denotes different
T878 3047-3050 NN denotes PKD
T879 3051-3057 NN denotes family
T880 3058-3065 NN denotes members
T881 3067-3075 JJ denotes Previous
T882 3076-3083 NN denotes studies
T883 3084-3088 VB denotes have
T884 3089-3094 VB denotes shown
T885 3095-3099 IN denotes that
T886 3100-3104 NN denotes PKDs
T887 3105-3108 VB denotes are
T888 3109-3122 JJ denotes indispensable
T889 3123-3126 IN denotes for
T890 3127-3131 NN denotes HDAC
T891 3132-3142 NN denotes regulation
T892 3143-3145 IN denotes in
T893 3146-3147 NN denotes B
T894 3148-3153 NN denotes cells
T895 3154-3155 -LRB- denotes [
T896 3155-3156 CD denotes 1
T897 3156-3157 -RRB- denotes ]
T898 3159-3165 RB denotes Herein
T899 3166-3168 PRP denotes we
T900 3169-3173 VB denotes show
T901 3174-3178 IN denotes that
T902 3179-3183 NN denotes PKDs
T903 3184-3187 VB denotes are
T904 3188-3192 RB denotes also
T905 3193-3206 JJ denotes indispensable
T906 3207-3210 IN denotes for
T907 3211-3216 NN denotes HSP27
T908 3217-3232 NN denotes phosphorylation
T909 3233-3235 IN denotes in
T910 3236-3237 NN denotes B
T911 3238-3243 NN denotes cells
T912 3245-3252 RB denotes However
T913 3252-3253 -COMMA- denotes ,
T914 3254-3262 JJ denotes PKD-null
T915 3263-3267 NN denotes DT40
T916 3268-3269 NN denotes B
T917 3270-3275 NN denotes cells
T918 3276-3279 VB denotes are
T919 3280-3286 JJ denotes viable
T920 3287-3290 CC denotes and
T921 3291-3302 VB denotes proliferate
T922 3303-3311 RB denotes normally
T923 3313-3321 RB denotes Moreover
T924 3321-3322 -COMMA- denotes ,
T925 3323-3327 NN denotes loss
T926 3328-3330 IN denotes of
T927 3331-3334 DT denotes the
T928 3335-3341 JJ denotes entire
T929 3342-3350 JJ denotes cellular
T930 3351-3355 NN denotes pool
T931 3356-3358 IN denotes of
T932 3359-3362 NN denotes PKD
T933 3363-3367 VB denotes does
T934 3368-3371 RB denotes not
T935 3372-3382 RB denotes critically
T936 3383-3389 VB denotes affect
T937 3390-3399 JJ denotes oxidative
T938 3400-3406 NN denotes stress
T939 3407-3416 NN denotes responses
T940 3417-3419 IN denotes in
T941 3420-3421 NN denotes B
T942 3422-3427 NN denotes cells
T943 3428-3431 CC denotes nor
T944 3432-3434 VB denotes do
T945 3435-3438 NN denotes PKD
T946 3439-3446 NN denotes kinases
T947 3447-3451 VB denotes play
T948 3452-3454 DT denotes an
T949 3455-3464 JJ denotes essential
T950 3465-3469 NN denotes role
T951 3470-3472 IN denotes in
T952 3473-3483 VB denotes regulating
T953 3484-3488 NN denotes NFκB
T954 3489-3504 JJ denotes transcriptional
T955 3505-3513 NN denotes activity
T956 3515-3523 RB denotes Together
T957 3523-3524 -COMMA- denotes ,
T958 3525-3530 DT denotes these
T959 3531-3539 NN denotes findings
T960 3540-3546 VB denotes reveal
T961 3547-3551 IN denotes that
T962 3552-3554 IN denotes in
T963 3555-3556 NN denotes B
T964 3557-3568 NN denotes lymphocytes
T965 3568-3569 -COMMA- denotes ,
T966 3570-3573 NN denotes PKD
T967 3574-3581 NN denotes kinases
T968 3582-3585 VB denotes are
T969 3586-3589 RB denotes not
T970 3590-3598 JJ denotes critical
T971 3599-3609 NN denotes regulators
T972 3610-3612 IN denotes of
T973 3613-3617 JJ denotes many
T974 3618-3620 IN denotes of
T975 3621-3624 DT denotes the
T976 3625-3633 JJ denotes cellular
T977 3634-3643 NN denotes processes
T978 3644-3654 RB denotes previously
T979 3655-3663 VB denotes ascribed
T980 3664-3666 TO denotes to
T981 3667-3671 PRP denotes them
T982 3672-3674 IN denotes in
T983 3675-3680 JJ denotes other
T984 3681-3689 JJ denotes cellular
T985 3690-3697 NN denotes systems
T1675 3736-3743 NN denotes culture
T1676 3743-3744 -COMMA- denotes ,
T1677 3745-3754 JJ denotes transient
T1678 3755-3768 NN denotes transfections
T1679 3769-3772 CC denotes and
T1680 3773-3777 NN denotes cell
T1681 3778-3789 NN denotes stimulation
T1682 3790-3793 DT denotes The
T1683 3794-3804 NN denotes generation
T1684 3804-3805 -COMMA- denotes ,
T1685 3806-3813 NN denotes culture
T1686 3814-3817 CC denotes and
T1687 3818-3828 NN denotes activation
T1688 3829-3831 IN denotes of
T1689 3832-3839 NN denotes PKD1−/−
T1690 3839-3840 -COMMA- denotes ,
T1691 3841-3848 NN denotes PKD3−/−
T1692 3849-3852 CC denotes and
T1693 3853-3862 NN denotes PKD1/3−/−
T1694 3863-3871 NN denotes knockout
T1695 3872-3876 NN denotes DT40
T1696 3877-3878 NN denotes B
T1697 3879-3883 NN denotes cell
T1698 3884-3889 NN denotes lines
T1699 3890-3894 VB denotes have
T1700 3895-3899 VB denotes been
T1701 3900-3909 VB denotes described
T1702 3910-3920 RB denotes previously
T1703 3921-3922 -LRB- denotes [
T1704 3922-3923 CD denotes 1
T1705 3923-3924 -RRB- denotes ]
T1706 3926-3931 NN denotes Cells
T1707 3932-3936 VB denotes were
T1708 3937-3942 VB denotes lysed
T1709 3943-3946 CC denotes and
T1710 3947-3954 NN denotes protein
T1711 3955-3963 NN denotes extracts
T1712 3964-3968 VB denotes were
T1713 3969-3977 VB denotes analysed
T1714 3978-3980 IN denotes in
T1715 3981-3988 JJ denotes Western
T1716 3989-3997 NN denotes blotting
T1717 3998-4009 NN denotes experiments
T1718 4010-4012 IN denotes as
T1719 4013-4023 RB denotes previously
T1720 4024-4033 VB denotes described
T1721 4034-4035 -LRB- denotes [
T1722 4035-4036 CD denotes 1
T1723 4036-4037 -RRB- denotes ]
T1724 4039-4054 NN denotes Chloramphenicol
T1725 4055-4061 NN denotes acetyl
T1726 4062-4073 NN denotes transferase
T1727 4074-4080 NN denotes assays
T1728 4081-4085 VB denotes have
T1729 4086-4090 VB denotes been
T1730 4091-4100 VB denotes described
T1731 4101-4111 RB denotes previously
T1732 4112-4113 -LRB- denotes [
T1733 4113-4115 CD denotes 29
T1734 4115-4116 -RRB- denotes ]
T1869 4129-4137 NN denotes staining
T1870 4138-4142 NN denotes DT40
T1871 4143-4144 NN denotes B
T1872 4145-4150 NN denotes cells
T1873 4151-4152 -LRB- denotes (
T1874 4152-4159 CD denotes 2 × 106
T1875 4160-4165 NN denotes cells
T1876 4166-4169 IN denotes per
T1877 4170-4175 NN denotes point
T1878 4175-4176 -RRB- denotes )
T1879 4177-4181 VB denotes were
T1880 4182-4193 VB denotes resuspended
T1881 4194-4196 IN denotes in
T1882 4197-4203 NN denotes 200 μl
T1883 4204-4210 NN denotes buffer
T1884 4211-4212 -LRB- denotes (
T1885 4212-4216 NN denotes RPMI
T1886 4217-4221 CD denotes 1640
T1887 4222-4227 NN denotes media
T1888 4227-4228 -COMMA- denotes ,
T1889 4229-4230 CD denotes 1
T1890 4230-4231 NN denotes %
T1891 4232-4238 JJ denotes foetal
T1892 4239-4243 NN denotes calf
T1893 4244-4249 NN denotes serum
T1894 4249-4250 -RRB- denotes )
T1895 4251-4261 VB denotes containing
T1896 4262-4274 JJ denotes anti-chicken
T1897 4275-4277 NN denotes M1
T1898 4278-4288 JJ denotes monoclonal
T1899 4289-4297 NN denotes antibody
T1900 4298-4308 VB denotes conjugated
T1901 4309-4311 TO denotes to
T1902 4312-4316 NN denotes FITC
T1903 4317-4320 IN denotes for
T1904 4321-4327 NN denotes 20 min
T1905 4328-4330 IN denotes on
T1906 4331-4334 NN denotes ice
T1907 4336-4339 DT denotes The
T1908 4340-4345 NN denotes cells
T1909 4346-4350 VB denotes were
T1910 4351-4357 VB denotes washed
T1911 4358-4363 RB denotes twice
T1912 4364-4367 CC denotes and
T1913 4368-4379 JJ denotes fluorescent
T1914 4380-4389 NN denotes intensity
T1915 4390-4393 VB denotes was
T1916 4394-4402 VB denotes analysed
T1917 4403-4405 IN denotes by
T1918 4406-4410 NN denotes flow
T1919 4411-4420 NN denotes cytometry
T1920 4422-4425 DT denotes All
T1921 4426-4433 NN denotes results
T1922 4434-4439 VB denotes shown
T1923 4440-4443 VB denotes are
T1924 4444-4458 JJ denotes representative
T1925 4459-4461 IN denotes of
T1926 4462-4464 IN denotes at
T1927 4465-4468 CD denotes two
T1928 4469-4471 TO denotes to
T1929 4472-4476 CD denotes four
T1930 4477-4488 JJ denotes independent
T1931 4489-4500 NN denotes experiments
T1932 4501-4507 IN denotes unless
T1933 4508-4517 RB denotes otherwise
T1934 4518-4527 VB denotes indicated
R10 T53 T52 arg2Of PKD,(
R100 T129 T127 arg1Of deacetylases,II
R101 T129 T128 arg1Of deacetylases,histone
R102 T129 T130 arg1Of deacetylases,in
R103 T132 T130 arg2Of lymphocytes,in
R104 T132 T131 arg1Of lymphocytes,B
R105 T135 T133 arg1Of Biol.,Mol.
R106 T135 T134 arg1Of Biol.,Cell
R107 T135 T136 arg1Of Biol.,26
R108 T135 T137 arg1Of Biol.,","
R109 T138 T137 arg2Of 1569–1577,","
R11 T54 T52 arg3Of ),(
R110 T138 T139 arg1Of 1569–1577,]
R111 T140 T142 arg1Of We,show
R112 T142 T141 arg1Of show,now
R113 T144 T145 arg1Of PKDs,are
R114 T144 T147 arg2Of PKDs,required
R115 T144 T149 arg1Of PKDs,regulate
R116 T147 T142 arg2Of required,show
R117 T147 T143 arg1Of required,that
R118 T147 T145 arg2Of required,are
R119 T147 T146 arg1Of required,also
R12 T55 T48 arg2Of enzymes,of
R120 T149 T147 arg3Of regulate,required
R121 T149 T148 arg1Of regulate,to
R122 T149 T152 arg1Of regulate,in
R123 T151 T149 arg2Of phosphorylation,regulate
R124 T151 T150 arg1Of phosphorylation,HSP27
R125 T154 T152 arg2Of B-cells,in
R126 T154 T153 arg1Of B-cells,DT40
R127 T158 T157 arg2Of contrast,in
R128 T158 T159 arg1Of contrast,to
R129 T161 T159 arg2Of observations,to
R13 T55 T51 arg1Of enzymes,D
R130 T161 T160 arg1Of observations,previous
R131 T161 T162 arg1Of observations,in
R132 T165 T162 arg2Of types,in
R133 T165 T163 arg1Of types,other
R134 T165 T164 arg1Of types,cell
R135 T168 T167 arg1Of enzymes,PKD
R136 T168 T169 arg1Of enzymes,do
R137 T168 T171 arg1Of enzymes,regulate
R138 T171 T155 arg1Of regulate,However
R139 T171 T156 arg1Of regulate,","
R14 T56 T45 arg1Of we,investigate
R140 T171 T157 arg1Of regulate,in
R141 T171 T166 arg1Of regulate,","
R142 T171 T169 arg2Of regulate,do
R143 T171 T170 arg1Of regulate,not
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
R144 T174 T171 arg2Of processes,regulate
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
R145 T174 T172 arg1Of processes,basic
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−/−
R146 T174 T173 arg1Of processes,cellular
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,[
R147 T174 T176 arg1Of processes,as
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
R148 T176 T175 arg1Of as,such
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 ],[
R149 T177 T178 arg1Of proliferation,or
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,[
R15 T56 T57 arg1Of we,generated
R150 T178 T182 arg1Of or,nor
R1500 T1733 T1732 arg2Of 29,[
R1501 T1734 T1732 arg3Of ],[
R151 T180 T178 arg2Of responses,or
R152 T180 T179 arg1Of responses,survival
R153 T182 T176 arg2Of nor,as
R154 T182 T181 arg1Of nor,","
R155 T185 T182 arg2Of activity,nor
R156 T185 T183 arg1Of activity,NFκB
R157 T185 T184 arg1Of activity,transcriptional
R158 T185 T186 arg1Of activity,downstream
R159 T186 T187 arg1Of downstream,of
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"
R1597 T1872 T1879 arg1Of cells,were
R1598 T1872 T1870 arg1Of cells,DT40
R1599 T1872 T1871 arg1Of cells,B
R16 T57 T44 modOf generated,To
R160 T192 T187 arg2Of receptor,of
R1600 T1872 T1873 arg1Of cells,(
R1601 T1872 T1880 arg2Of cells,resuspended
R1602 T1875 T1874 arg1Of cells,2 × 106
R1603 T1875 T1876 arg1Of cells,per
R1604 T1875 T1873 arg2Of cells,(
R1605 T1877 T1876 arg2Of point,per
R1606 T1878 T1873 arg3Of ),(
R1607 T1880 T1879 arg2Of resuspended,were
R1608 T1880 T1881 arg1Of resuspended,in
R1609 T1883 T1881 arg2Of buffer,in
R161 T192 T188 arg1Of receptor,the
R1610 T1883 T1882 arg1Of buffer,200 μl
R1611 T1883 T1884 arg1Of buffer,(
R1612 T1883 T1895 arg1Of buffer,containing
R1613 T1887 T1884 arg2Of media,(
R1614 T1887 T1885 arg1Of media,RPMI
R1615 T1887 T1886 arg1Of media,1640
R1616 T1887 T1888 arg1Of media,","
R1617 T1889 T1890 arg1Of 1,%
R1618 T1893 T1888 arg2Of serum,","
R1619 T1893 T1889 arg1Of serum,1
R162 T192 T189 arg1Of receptor,B
R1620 T1893 T1891 arg1Of serum,foetal
R1621 T1893 T1892 arg1Of serum,calf
R1622 T1894 T1884 arg3Of ),(
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
R163 T192 T190 arg1Of receptor,cell
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
R164 T192 T191 arg1Of receptor,antigen
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
R165 T195 T196 arg1Of PKDs,have
R1650 T1921 T1923 arg1Of results,are
R1651 T1921 T1924 arg1Of results,representative
R1652 T1923 T1932 arg1Of are,unless
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
R166 T196 T193 arg1Of have,Thus
R1660 T1931 T1930 arg1Of experiments,independent
R1661 T1934 T1932 arg2Of indicated,unless
R1662 T1934 T1933 arg1Of indicated,otherwise
R167 T196 T194 arg1Of have,","
R168 T196 T200 arg1Of have,in
R169 T199 T196 arg2Of role,have
R17 T63 T57 arg2Of line,generated
R170 T199 T197 arg1Of role,a
R171 T199 T198 arg1Of role,selective
R172 T203 T200 arg2Of biology,in
R173 T203 T201 arg1Of biology,DT40
R174 T203 T202 arg1Of biology,B-cell
R18 T63 T58 arg1Of line,a
R19 T63 T59 arg1Of line,PKD-null
R20 T63 T60 arg1Of line,DT40
R21 T63 T61 arg1Of line,B-lymphocyte
R22 T63 T62 arg1Of line,cell
R23 T65 T66 arg1Of we,have
R24 T65 T67 arg1Of we,shown
R25 T67 T64 arg1Of shown,Previously
R26 T67 T66 arg2Of shown,have
R27 T69 T70 arg1Of PKDs,have
R28 T70 T67 arg2Of have,shown
R29 T70 T68 arg1Of have,that
R30 T70 T74 arg1Of have,in
R31 T73 T70 arg2Of role,have
R32 T73 T71 arg1Of role,an
R33 T73 T72 arg1Of role,essential
R34 T75 T74 arg2Of regulating,in
R35 T75 T80 arg1Of regulating,in
R36 T79 T75 arg2Of deacetylases,regulating
R37 T79 T76 arg1Of deacetylases,class
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
R38 T79 T77 arg1Of deacetylases,II
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
R39 T79 T78 arg1Of deacetylases,histone
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
R40 T82 T80 arg2Of B-cells,in
R41 T82 T81 arg1Of B-cells,DT40
R42 T84 T85 arg1Of Matthews,","
R43 T85 T87 arg1Of ",",","
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
R44 T86 T85 arg2Of S.A.,","
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
R45 T87 T89 arg1Of ",",","
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
R46 T88 T87 arg2Of Liu,","
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
R47 T89 T91 arg1Of ",",","
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
R48 T90 T89 arg2Of P.,","
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
R49 T91 T93 arg1Of ",",","
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
R50 T92 T91 arg2Of Spitaler,","
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
R51 T93 T95 arg1Of ",",","
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
R52 T94 T93 arg2Of M.,","
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
R53 T95 T105 arg1Of ",",","
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
R54 T96 T97 arg1Of Olson,","
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
R55 T97 T99 arg1Of ",",","
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
R56 T98 T97 arg2Of E.N.,","
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,[
R57 T99 T101 arg1Of ",",","
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
R58 T100 T99 arg2Of McKinsey,","
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,[
R59 T101 T103 arg1Of ",",","
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
R60 T102 T101 arg2Of T.A.,","
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
R61 T103 T95 arg2Of ",",","
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,","
R62 T104 T103 arg2Of Cantrell,","
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 ],[
R63 T105 T107 arg1Of ",",and
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
R64 T106 T105 arg2Of D.A.,","
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
R65 T108 T109 arg1Of Scharenberg,","
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",[
R66 T109 T107 arg2Of ",",and
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,[
R67 T110 T109 arg2Of A.M.,","
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,[
R68 T112 T111 arg2Of 2006,(
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
R69 T113 T111 arg3Of ),(
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
R7 T51 T49 arg1Of D,protein
R70 T115 T80 arg3Of role,in
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
R71 T115 T83 arg1Of role,[
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
R72 T115 T84 arg1Of role,Matthews
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
R73 T115 T86 arg1Of role,S.A.
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
R74 T115 T88 arg1Of role,Liu
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,[
R75 T115 T90 arg1Of role,P.
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
R76 T115 T92 arg1Of role,Spitaler
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
R77 T115 T94 arg1Of role,M.
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
R78 T115 T96 arg1Of role,Olson
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
R79 T115 T98 arg1Of role,E.N.
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
R8 T51 T50 arg1Of D,kinase
R80 T115 T100 arg1Of role,McKinsey
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
R81 T115 T102 arg1Of role,T.A.
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
R82 T115 T104 arg1Of role,Cantrell
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
R83 T115 T106 arg1Of role,D.A.
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
R84 T115 T108 arg1Of role,Scharenberg
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
R85 T115 T110 arg1Of role,A.M.
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
R86 T115 T111 arg1Of role,(
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
R87 T115 T114 arg1Of role,Essential
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
R88 T115 T116 arg1Of role,for
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
R89 T115 T122 arg1Of role,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
R9 T51 T52 arg1Of D,(
R90 T121 T116 arg2Of kinases,for
R91 T121 T117 arg1Of kinases,protein
R92 T121 T118 arg1Of kinases,kinase
R93 T121 T119 arg1Of kinases,D
R94 T121 T120 arg1Of kinases,family
R95 T124 T122 arg2Of regulation,in
R96 T124 T123 arg1Of regulation,the
R97 T124 T125 arg1Of regulation,of
R98 T129 T125 arg2Of deacetylases,of
R99 T129 T126 arg1Of deacetylases,class