0 18 Cell-type-specific cell-type-specific JJ 19 29 regulation regulation NN 30 32 of of IN 33 36 the the DT 37 42 human human JJ 43 48 tumor tumor NN 49 57 necrosis necrosis NN 58 64 factor factor NN 65 70 alpha alpha NN 71 75 gene gene NN 76 78 in in IN 79 80 B B NNP 81 86 cells cell NNS 87 90 and and CC 91 92 T t NN 93 98 cells cell NNS 99 101 by by IN 102 107 NFATp NFATp NNP 108 111 and and CC 112 121 ATF-2/JUN ATF-2/JUN NNP 121 122 . . . 124 127 The the DT 128 133 human human JJ 134 139 tumor tumor NN 140 148 necrosis necrosis NN 149 155 factor factor NN 156 161 alpha alpha NN 162 163 ( ( ( 163 172 TNF-alpha TNF-alpha NNP 172 173 ) ) ) 174 178 gene gene NN 179 181 is be VBZ 182 185 one one CD 186 188 of of IN 189 192 the the DT 193 201 earliest early JJS 202 207 genes gene NNS 208 219 transcribed transcribe VBN 220 225 after after IN 226 229 the the DT 230 241 stimulation stimulation NN 242 244 of of IN 245 246 a a DT 247 248 B B NNP 249 253 cell cell NN 254 261 through through IN 262 265 its its PRP$ 266 273 antigen antigen NN 274 282 receptor receptor NN 283 285 or or CC 286 289 via via IN 290 293 the the DT 294 299 CD-40 cd-40 NN 300 307 pathway pathway NN 307 308 . . . 309 311 In in IN 312 316 both both DT 317 322 cases case NNS 322 323 , , , 324 333 induction induction NN 334 336 of of IN 337 346 TNF-alpha TNF-alpha NNP 347 351 gene gene NN 352 365 transcription transcription NN 366 369 can can MD 370 372 be be VB 373 380 blocked block VBN 381 383 by by IN 384 387 the the DT 388 406 immunosuppressants immunosuppressant NNS 407 418 cyclosporin cyclosporin NN 419 420 A A NNP 421 424 and and CC 425 430 FK506 FK506 NNP 430 431 , , , 432 437 which which WDT 438 447 suggested suggest VBD 448 449 a a DT 450 454 role role NN 455 458 for for IN 459 462 the the DT 463 467 NFAT nfat NN 468 474 family family NN 475 477 of of IN 478 486 proteins protein NNS 487 489 in in IN 490 493 the the DT 494 504 regulation regulation NN 505 507 of of IN 508 511 the the DT 512 516 gene gene NN 517 519 in in IN 520 521 B B NNP 522 527 cells cell NNS 527 528 . . . 529 540 Furthermore furthermore RB 540 541 , , , 542 544 in in IN 545 546 T t NN 547 552 cells cell NNS 552 553 , , , 554 557 two two CD 558 567 molecules molecule NNS 568 570 of of IN 571 576 NFATp NFATp NNP 577 581 bind bind VBP 582 584 to to TO 585 588 the the DT 589 598 TNF-alpha TNF-alpha NNP 599 607 promoter promoter NN 608 615 element element NN 616 621 kappa kappa NN 622 623 3 3 CD 624 626 in in IN 627 638 association association NN 639 643 with with IN 644 649 ATF-2 ATF-2 NNP 650 653 and and CC 654 657 Jun Jun NNP 658 666 proteins protein NNS 667 672 bound bind VBD 673 675 to to TO 676 678 an an DT 679 690 immediately immediately RB 691 699 adjacent adjacent JJ 700 706 cyclic cyclic JJ 707 710 AMP AMP NNP 711 719 response response NN 720 727 element element NN 728 729 ( ( ( 729 732 CRE CRE NNP 732 733 ) ) ) 734 738 site site NN 738 739 . . . 740 744 Here here RB 744 745 , , , 746 751 using use VBG 752 755 the the DT 756 762 murine murine JJ 763 769 B-cell b-cell NN 770 778 lymphoma lymphoma NN 779 783 cell cell NN 784 788 line line NN 789 792 A20 a20 NN 792 793 , , , 794 796 we we PRP 797 801 show show VBP 802 806 that that IN 807 810 the the DT 811 820 TNF-alpha TNF-alpha NNP 821 825 gene gene NN 826 828 is be VBZ 829 838 regulated regulate VBN 839 841 in in IN 842 843 a a DT 844 862 cell-type-specific cell-type-specific JJ 863 869 manner manner NN 869 870 . . . 871 873 In in IN 874 877 A20 A20 NNP 878 879 B B NNP 880 885 cells cell NNS 885 886 , , , 887 890 the the DT 891 900 TNF-alpha TNF-alpha NNP 901 905 gene gene NN 906 908 is be VBZ 909 912 not not RB 913 922 regulated regulate VBN 923 925 by by IN 926 931 NFATp NFATp NNP 932 937 bound bind VBD 938 940 to to TO 941 944 the the DT 945 950 kappa kappa NN 951 952 3 3 CD 953 960 element element NN 960 961 . . . 962 969 Instead instead RB 969 970 , , , 971 976 ATF-2 ATF-2 NNP 977 980 and and CC 981 984 Jun Jun NNP 985 993 proteins protein NNS 994 998 bind bind VBP 999 1001 to to TO 1002 1005 the the DT 1006 1015 composite composite JJ 1016 1021 kappa kappa NN 1022 1027 3/CRE 3/cre NN 1028 1032 site site NN 1033 1036 and and CC 1037 1042 NFATp nfatp NN 1043 1048 binds bind VBZ 1049 1051 to to TO 1052 1053 a a DT 1054 1059 newly newly RB 1060 1070 identified identify VBN 1071 1077 second second JJ 1078 1082 NFAT NFAT NNP 1083 1087 site site NN 1088 1096 centered centered JJ 1097 1099 at at IN 1100 1103 -76 -76 CD 1104 1115 nucleotides nucleotide NNS 1116 1124 relative relative JJ 1125 1127 to to TO 1128 1131 the the DT 1132 1141 TNF-alpha TNF-alpha NNP 1142 1155 transcription transcription NN 1156 1161 start start NN 1162 1166 site site NN 1166 1167 . . . 1168 1172 This this DT 1173 1176 new new JJ 1177 1181 site site NN 1182 1187 plays play VBZ 1188 1189 a a DT 1190 1198 critical critical JJ 1199 1203 role role NN 1204 1206 in in IN 1207 1210 the the DT 1211 1227 calcium-mediated calcium-mediated JJ 1227 1228 , , , 1229 1240 cyclosporin cyclosporin NN 1241 1252 A-sensitive a-sensitive JJ 1253 1262 induction induction NN 1263 1265 of of IN 1266 1275 TNF-alpha TNF-alpha NNP 1276 1278 in in IN 1279 1283 both both CC 1284 1287 A20 A20 NNP 1288 1289 B B NNP 1290 1295 cells cell NNS 1296 1299 and and CC 1300 1304 Ar-5 ar-5 NN 1305 1310 cells cell NNS 1310 1311 . . . 1312 1322 Consistent consistent JJ 1323 1327 with with IN 1328 1333 these these DT 1334 1341 results result NNS 1341 1342 , , , 1343 1355 quantitative quantitative JJ 1356 1361 DNase dnase NN 1362 1374 footprinting footprinting NN 1375 1377 of of IN 1378 1381 the the DT 1382 1391 TNF-alpha TNF-alpha NNP 1392 1400 promoter promoter NN 1401 1406 using use VBG 1407 1417 increasing increase VBG 1418 1425 amounts amount NNS 1426 1428 of of IN 1429 1440 recombinant recombinant JJ 1441 1446 NFATp nfatp NN 1447 1459 demonstrated demonstrate VBD 1460 1464 that that IN 1465 1468 the the DT 1469 1472 -76 -76 CD 1473 1477 site site NN 1478 1483 binds bind VBZ 1484 1486 to to TO 1487 1492 NFATp nfatp NN 1493 1497 with with IN 1498 1499 a a DT 1500 1506 higher high JJR 1507 1515 affinity affinity NN 1516 1520 than than IN 1521 1524 the the DT 1525 1530 kappa kappa NN 1531 1532 3 3 CD 1533 1537 site site NN 1537 1538 . . . 1539 1542 Two two CD 1543 1548 other other JJ 1549 1559 previously previously RB 1560 1572 unrecognized unrecognized JJ 1573 1586 NFATp-binding nfatp-binding JJ 1587 1592 sites site NNS 1593 1595 in in IN 1596 1599 the the DT 1600 1608 proximal proximal JJ 1609 1618 TNF-alpha TNF-alpha NNP 1619 1627 promoter promoter NN 1628 1632 were be VBD 1633 1637 also also RB 1638 1648 identified identify VBN 1649 1651 by by IN 1652 1656 this this DT 1657 1665 analysis analysis NN 1665 1666 . . . 1667 1671 Thus thus RB 1671 1672 , , , 1673 1680 through through IN 1681 1684 the the DT 1685 1697 differential differential JJ 1698 1701 use use NN 1702 1704 of of IN 1705 1708 the the DT 1709 1713 same same JJ 1714 1722 promoter promoter NN 1723 1730 element element NN 1730 1731 , , , 1732 1735 the the DT 1736 1745 composite composite JJ 1746 1751 kappa kappa NN 1752 1757 3/CRE 3/cre NN 1758 1762 site site NN 1762 1763 , , , 1764 1767 the the DT 1768 1777 TNF-alpha TNF-alpha NNP 1778 1782 gene gene NN 1783 1785 is be VBZ 1786 1795 regulated regulate VBN 1796 1798 in in IN 1799 1800 a a DT 1801 1819 cell-type-specific cell-type-specific JJ 1820 1826 manner manner NN 1827 1829 in in IN 1830 1838 response response NN 1839 1841 to to TO 1842 1845 the the DT 1846 1850 same same JJ 1851 1864 extracellular extracellular JJ 1865 1871 signal signal NN 1871 1872 . . .