0 19 Anti-immunoglobulin anti-immunoglobulin JJ 20 21 M M NNP 22 31 activates activate VBZ 32 39 nuclear nuclear JJ 40 68 calcium/calmodulin-dependent calcium/calmodulin-dependent JJ 69 76 protein protein NN 77 83 kinase kinase NNP 84 86 II ii CD 87 89 in in IN 90 95 human human JJ 96 97 B B NNP 98 109 lymphocytes lymphocyte NNS 109 110 . . . 112 114 We we PRP 115 118 and and CC 119 125 others other NNS 126 130 have have VBP 131 141 previously previously RB 142 147 shown show VBN 148 152 that that IN 153 156 the the DT 157 164 nuclear nuclear JJ 165 172 protein protein NN 172 173 , , , 174 179 Ets-1 Ets-1 NNP 179 180 , , , 181 183 is be VBZ 184 198 phosphorylated phosphorylate VBN 199 201 in in IN 202 203 a a DT 204 221 calcium-dependent calcium-dependent JJ 222 228 manner manner NN 229 234 after after IN 235 243 ligation ligation NN 244 246 of of IN 247 261 immunoglobulin immunoglobulin NN 262 263 ( ( ( 263 265 Ig Ig NNP 265 266 ) ) ) 267 268 M M NNP 269 271 on on IN 272 273 B B NNP 274 285 lymphocytes lymphocyte NNS 285 286 . . . 287 289 As as IN 290 294 this this DT 295 310 phosphorylation phosphorylation NN 311 314 was be VBD 315 326 independent independent JJ 327 329 of of IN 330 337 protein protein NN 338 344 kinase kinase NNP 345 346 C C NNP 347 355 activity activity NN 355 356 , , , 357 359 we we PRP 360 366 tested test VBD 367 374 whether whether IN 375 376 a a DT 377 405 calcium/calmodulin-dependent calcium/calmodulin-dependent JJ 406 413 protein protein NN 414 420 kinase kinase NNP 421 422 ( ( ( 422 425 CaM CaM NNP 426 432 kinase kinase NNP 432 433 ) ) ) 434 439 might might MD 440 453 phosphorylate phosphorylate VB 454 457 the the DT 458 463 Ets-1 ets-1 NN 464 471 protein protein NN 472 477 after after IN 478 487 elevation elevation NN 488 490 of of IN 491 504 intracellular intracellular JJ 505 509 free free JJ 510 517 calcium calcium NN 518 532 concentrations concentration NNS 532 533 . . . 534 537 The the DT 538 554 dephosphorylated dephosphorylate VBN 555 559 form form NN 560 562 of of IN 563 568 Ets-1 Ets-1 NNP 569 572 has have VBZ 573 577 been be VBN 578 583 shown show VBN 584 586 to to TO 587 591 bind bind VB 592 594 to to TO 595 604 chromatin chromatin NN 604 605 , , , 606 616 suggesting suggest VBG 617 621 that that IN 622 625 the the DT 626 635 operative operative JJ 636 642 kinase kinase NNP 643 649 should should MD 650 652 be be VB 653 663 detectable detectable JJ 664 666 in in IN 667 670 the the DT 671 678 nucleus nucleus NN 678 679 . . . 680 682 We we PRP 683 691 prepared prepare VBD 692 699 nuclear nuclear JJ 700 708 extracts extract NNS 709 713 from from IN 714 717 two two CD 718 723 human human JJ 724 725 B B NNP 726 730 cell cell NN 731 736 lines line NNS 737 739 in in IN 740 745 which which WDT 746 755 increased increase VBD 756 769 intracellular intracellular JJ 770 774 free free JJ 775 782 calcium calcium NN 783 789 levels level NNS 790 800 correlated correlate VBD 801 805 with with IN 806 815 increased increase VBN 816 831 phosphorylation phosphorylation NN 832 834 of of IN 835 838 the the DT 839 844 Ets-1 ets-1 NN 845 852 protein protein NN 852 853 . . . 854 862 Activity activity NN 863 865 of of IN 866 869 the the DT 870 873 CaM CaM NNP 874 881 kinases kinase NNS 882 885 was be VBD 886 896 determined determine VBN 897 902 using use VBG 903 904 a a DT 905 914 synthetic synthetic JJ 915 922 peptide peptide NN 923 932 substrate substrate NN 933 937 both both CC 938 940 in in IN 941 944 the the DT 945 952 absence absence NN 953 956 and and CC 957 965 presence presence NN 966 968 of of IN 969 971 an an DT 972 981 inhibitor inhibitor NN 982 990 specific specific JJ 991 994 for for IN 995 998 the the DT 999 1002 CaM CaM NNP 1003 1009 kinase kinase NNP 1010 1016 family family NN 1016 1017 , , , 1018 1023 KN-62 kn-62 NN 1023 1024 . . . 1025 1036 Stimulation stimulation NN 1037 1039 of of IN 1040 1045 cells cell NNS 1046 1050 with with IN 1051 1059 anti-IgM anti-igm JJ 1060 1063 led lead VBD 1064 1066 to to TO 1067 1076 increased increase VBN 1077 1085 activity activity NN 1086 1088 of of IN 1089 1090 a a DT 1091 1098 nuclear nuclear JJ 1099 1105 kinase kinase NNP 1106 1110 that that WDT 1111 1116 could could MD 1117 1130 phosphorylate phosphorylate VB 1131 1134 the the DT 1135 1142 peptide peptide NN 1142 1143 , , , 1144 1147 and and CC 1148 1152 this this DT 1153 1161 activity activity NN 1162 1165 was be VBD 1166 1173 reduced reduce VBN 1174 1176 by by IN 1177 1179 10 10 CD 1180 1186 microM microm NN 1187 1192 KN-62 kn-62 NN 1192 1193 . . . 1194 1200 Kinase Kinase NNP 1201 1209 activity activity NN 1210 1213 was be VBD 1214 1221 reduced reduce VBN 1222 1224 in in IN 1225 1232 lysates lysate NNS 1233 1244 preadsorbed preadsorbe VBN 1245 1250 using use VBG 1251 1253 an an DT 1254 1262 antibody antibody NN 1263 1271 specific specific JJ 1272 1275 for for IN 1276 1279 CaM CaM NNP 1280 1286 kinase kinase NNP 1287 1289 II ii CD 1289 1290 . . . 1291 1306 Two-dimensional two-dimensional JJ 1307 1321 phosphopeptide phosphopeptide NN 1322 1326 maps map NNS 1327 1329 of of IN 1330 1333 the the DT 1334 1339 Ets-1 ets-1 NN 1340 1347 protein protein NN 1348 1352 from from IN 1353 1358 cells cell NNS 1359 1368 incubated incubate VBN 1369 1373 with with IN 1374 1383 ionomycin ionomycin NN 1384 1386 or or CC 1387 1395 anti-IgM anti-IgM NNP 1396 1405 contained contain VBD 1406 1409 two two CD 1410 1416 unique unique JJ 1417 1432 phosphopeptides phosphopeptide NNS 1433 1437 that that WDT 1438 1442 were be VBD 1443 1449 absent absent JJ 1450 1452 in in IN 1453 1462 untreated untreated JJ 1463 1468 cells cell NNS 1468 1469 . . . 1470 1480 Incubation incubation NN 1481 1483 of of IN 1484 1492 isolated isolate VBN 1493 1498 Ets-1 ets-1 NN 1499 1506 protein protein NN 1507 1511 with with IN 1512 1520 purified purify VBN 1521 1524 CaM CaM NNP 1525 1531 kinase kinase NNP 1532 1534 II ii CD 1535 1543 produced produce VBD 1544 1559 phosphorylation phosphorylation NN 1560 1562 of of IN 1563 1571 peptides peptide NNS 1572 1576 that that WDT 1577 1585 migrated migrate VBD 1586 1597 identically identically RB 1598 1600 to to TO 1601 1606 those those DT 1607 1612 found find VBN 1613 1615 in in IN 1616 1621 cells cell NNS 1622 1631 incubated incubate VBN 1632 1636 with with IN 1637 1643 either either CC 1644 1652 anti-IgM anti-igm JJ 1653 1655 or or CC 1656 1665 ionomycin ionomycin NN 1665 1666 . . . 1667 1672 These these DT 1673 1677 data datum NNS 1678 1685 suggest suggest VBP 1686 1687 a a DT 1688 1693 model model NN 1694 1696 of of IN 1697 1703 signal signal NN 1704 1716 transduction transduction NN 1717 1719 by by IN 1720 1723 the the DT 1724 1731 antigen antigen NN 1732 1740 receptor receptor NN 1741 1743 on on IN 1744 1745 B B NNP 1746 1757 lymphocytes lymphocyte NNS 1758 1760 in in IN 1761 1766 which which WDT 1767 1776 increased increase VBD 1777 1790 intracellular intracellular JJ 1791 1795 free free JJ 1796 1803 calcium calcium NN 1804 1807 can can MD 1808 1815 rapidly rapidly RB 1816 1824 activate activate VB 1825 1832 nuclear nuclear JJ 1833 1836 CaM CaM NNP 1837 1843 kinase kinase NNP 1844 1846 II ii CD 1846 1847 , , , 1848 1859 potentially potentially RB 1860 1869 resulting result VBG 1870 1872 in in IN 1873 1888 phosphorylation phosphorylation NN 1889 1892 and and CC 1893 1903 regulation regulation NN 1904 1906 of of IN 1907 1918 DNA-binding dna-binding JJ 1919 1927 proteins protein NNS 1927 1928 . . .