0 1 A a DT 2 9 nuclear nuclear JJ 10 16 factor factor NN 17 23 NF-GM2 nf-gm2 NN 24 28 that that WDT 29 38 interacts interact VBZ 39 43 with with IN 44 45 a a DT 46 56 regulatory regulatory JJ 57 63 region region NN 64 66 of of IN 67 70 the the DT 71 77 GM-CSF GM-CSF NNP 78 82 gene gene NN 83 92 essential essential JJ 93 96 for for IN 97 100 its its PRP$ 101 110 induction induction NN 111 113 in in IN 114 123 responses response NNS 124 126 to to TO 127 133 T-cell t-cell NN 134 144 activation activation NN 144 145 : : : 146 158 purification purification NN 159 163 from from IN 164 169 human human JJ 170 176 T-cell t-cell NN 177 185 leukemia leukemia NN 186 190 line line NN 191 197 Jurkat jurkat NN 198 203 cells cell NNS 204 207 and and CC 208 218 similarity similarity NN 219 221 to to TO 222 230 NF-kappa NF-kappa NNP 231 233 B. B. NNP 235 245 Activation Activation NNP 246 248 of of IN 249 250 T t NN 251 256 cells cell NNS 257 259 by by IN 260 267 antigen antigen NN 267 268 , , , 269 275 lectin lectin NN 275 276 , , , 277 279 or or CC 280 281 a a DT 282 293 combination combination NN 294 296 of of IN 297 317 phorbol-12-myristate phorbol-12-myristate NN 318 325 acetate acetate NN 326 327 ( ( ( 327 330 PMA PMA NNP 330 331 ) ) ) 332 335 and and CC 336 343 calcium calcium NN 344 353 ionophore ionophore NN 354 355 ( ( ( 355 361 A23187 a23187 NN 361 362 ) ) ) 363 368 leads lead VBZ 369 371 to to TO 372 375 the the DT 376 385 induction induction NN 386 388 of of IN 389 394 genes gene NNS 395 398 for for IN 399 400 a a DT 401 404 set set NN 405 407 of of IN 408 419 lymphokines lymphokine NNS 419 420 , , , 421 430 including include VBG 431 453 granulocyte-macrophage granulocyte-macrophage JJ 454 472 colony-stimulating colony-stimulating JJ 473 479 factor factor NN 480 481 ( ( ( 481 487 GM-CSF GM-CSF NNP 487 488 ) ) ) 488 489 . . . 490 492 We we PRP 493 505 demonstrated demonstrate VBD 506 508 in in IN 509 516 earlier early JJR 517 524 studies study NNS 525 529 that that IN 530 533 the the DT 534 542 upstream upstream JJ 543 549 region region NN 550 552 of of IN 553 556 the the DT 557 562 mouse mouse NN 563 569 GM-CSF GM-CSF NNP 570 578 promoter promoter NN 579 581 at at IN 582 591 positions position NNS 592 599 between between IN 600 603 -95 -95 CD 604 607 and and CC 608 611 -73 -73 CD 612 614 is be VBZ 615 624 essential essential JJ 625 628 for for IN 629 644 transcriptional transcriptional JJ 645 655 activation activation NN 656 658 in in IN 659 667 response response NN 668 670 to to TO 671 681 PMA/A23187 pma/a23187 NN 681 682 . . . 683 687 This this DT 688 694 region region NN 695 703 contains contain VBZ 704 707 two two CD 708 719 DNA-binding dna-binding JJ 720 726 motifs motif NNS 726 727 , , , 728 731 GM2 GM2 NNP 732 735 and and CC 736 742 GC-box GC-box NNP 742 743 . . . 744 747 The the DT 748 751 GM2 gm2 NN 752 760 sequence sequence NN 761 762 ( ( ( 762 772 GGTAGTTCCC GGTAGTTCCC NNP 772 773 ) ) ) 774 776 is be VBZ 777 787 recognized recognize VBN 788 790 by by IN 791 793 an an DT 794 803 inducible inducible JJ 804 810 factor factor NN 811 817 NF-GM2 nf-gm2 NN 817 818 ; ; : 819 822 the the DT 823 828 other other JJ 829 830 ( ( ( 830 836 CCGCCC CCGCCC NNP 836 837 ) ) ) 838 840 by by IN 841 853 constitutive constitutive JJ 854 861 factors factor NNS 862 864 A1 a1 NN 864 865 , , , 866 868 A2 A2 NNP 868 869 , , , 870 873 and and CC 874 876 B. B. NNP 877 879 To to TO 880 889 elucidate elucidate VB 890 893 the the DT 894 903 mechanism mechanism NN 904 906 of of IN 907 913 GM-CSF GM-CSF NNP 914 918 gene gene NN 919 929 activation activation NN 929 930 , , , 931 933 we we PRP 934 938 have have VBP 939 947 purified purify VBN 948 951 the the DT 952 961 inducible inducible JJ 962 968 factor factor NN 969 975 NF-GM2 nf-gm2 NN 976 980 from from IN 981 984 the the DT 985 992 nuclear nuclear JJ 993 1000 extract extract NN 1001 1003 of of IN 1004 1014 stimulated stimulate VBN 1015 1021 Jurkat jurkat NN 1022 1027 cells cell NNS 1028 1030 on on IN 1031 1034 the the DT 1035 1040 basis basis NN 1041 1043 of of IN 1044 1052 specific specific JJ 1053 1064 DNA-binding dna-binding JJ 1065 1073 activity activity NN 1073 1074 . . . 1075 1078 The the DT 1079 1087 purified purify VBN 1088 1094 NF-GM2 nf-gm2 NN 1095 1103 consists consist VBZ 1104 1106 of of IN 1107 1109 50 50 CD 1110 1111 ( ( ( 1111 1114 p50 p50 NN 1114 1115 ) ) ) 1116 1119 and and CC 1120 1122 65 65 CD 1123 1126 kDa kda NN 1127 1128 ( ( ( 1128 1131 p65 p65 NN 1131 1132 ) ) ) 1133 1145 polypeptides polypeptide NNS 1146 1149 and and CC 1150 1153 has have VBZ 1154 1155 a a DT 1156 1163 binding binding NN 1164 1172 activity activity NN 1173 1181 specific specific JJ 1182 1185 for for IN 1186 1190 both both CC 1191 1194 the the DT 1195 1201 GM-CSF GM-CSF NNP 1202 1205 and and CC 1206 1220 immunoglobulin immunoglobulin NN 1221 1226 kappa kappa NN 1227 1228 ( ( ( 1228 1238 GGAAAGTCCC GGAAAGTCCC NNP 1238 1239 ) ) ) 1240 1249 enhancers enhancer NNS 1249 1250 . . . 1251 1270 Electrophoretically electrophoretically RB 1271 1279 purified purify VBN 1280 1283 p50 p50 NN 1284 1289 alone alone RB 1290 1293 can can MD 1294 1298 form form VB 1299 1300 a a DT 1301 1312 protein-DNA protein-dna JJ 1313 1320 complex complex NN 1320 1321 , , , 1322 1325 but but CC 1326 1328 in in IN 1329 1332 the the DT 1333 1340 mixture mixture NN 1340 1341 , , , 1342 1345 p50 p50 NN 1346 1356 associates associate VBZ 1357 1371 preferentially preferentially RB 1372 1376 with with IN 1377 1380 p65 p65 NN 1381 1383 to to TO 1384 1388 form form VB 1389 1392 the the DT 1393 1399 NF-GM2 nf-gm2 NN 1400 1407 complex complex NN 1407 1408 . . . 1409 1411 In in IN 1412 1420 addition addition NN 1420 1421 , , , 1422 1425 p65 p65 NN 1426 1430 gave give VBD 1431 1434 per per FW 1435 1437 se se FW 1437 1438 , , , 1439 1443 with with IN 1444 1447 low low JJ 1448 1456 affinity affinity NN 1456 1457 , , , 1458 1459 a a DT 1460 1471 protein-DNA protein-dna JJ 1472 1479 complex complex NN 1480 1484 that that WDT 1485 1493 migrated migrate VBD 1494 1498 more more RBR 1499 1505 slowly slowly RB 1506 1510 than than IN 1511 1517 native native JJ 1518 1524 NF-GM2 nf-gm2 NN 1525 1532 complex complex NN 1532 1533 . . . 1534 1545 Furthermore furthermore RB 1545 1546 , , , 1547 1549 an an DT 1550 1559 antiserum antiserum NN 1560 1567 against against IN 1568 1572 KBF1 kbf1 NN 1573 1574 ( ( ( 1574 1583 identical identical JJ 1584 1586 to to TO 1587 1589 50 50 CD 1590 1593 kDa kda NN 1594 1602 NF-kappa NF-kappa NNP 1603 1604 B B NNP 1605 1612 protein protein NN 1612 1613 ) ) ) 1614 1621 reacted react VBD 1622 1626 with with IN 1627 1630 the the DT 1631 1634 p50 p50 NN 1635 1637 of of IN 1638 1644 NF-GM2 nf-gm2 NN 1644 1645 , , , 1646 1656 indicating indicate VBG 1657 1661 that that IN 1662 1665 the the DT 1666 1672 NF-GM2 nf-gm2 NN 1673 1684 polypeptide polypeptide NN 1685 1691 cannot cannot MD 1692 1694 be be VB 1695 1710 immunologically immunologically RB 1711 1725 differentiated differentiate VBN 1726 1730 from from IN 1731 1734 the the DT 1735 1737 50 50 CD 1738 1741 kDa kda NN 1742 1749 subunit subunit NN 1750 1752 of of IN 1753 1761 NF-kappa NF-kappa NNP 1762 1764 B. B. NNP 1765 1768 The the DT 1769 1777 purified purify VBN 1778 1784 NF-GM2 nf-gm2 NN 1785 1794 activated activate VBN 1795 1797 in in FW 1798 1803 vitro vitro FW 1804 1817 transcription transcription NN 1818 1822 from from IN 1823 1826 the the DT 1827 1832 kappa kappa NN 1833 1834 B B NNP 1835 1843 enhancer enhancer NN 1843 1844 , , , 1845 1850 while while IN 1851 1853 it it PRP 1854 1860 failed fail VBD 1861 1863 to to TO 1864 1873 stimulate stimulate VB 1874 1887 transcription transcription NN 1888 1892 from from IN 1893 1896 the the DT 1897 1903 GM-CSF GM-CSF NNP 1904 1912 promoter promoter NN 1913 1922 harboring harbor VBG 1923 1926 the the DT 1927 1930 GM2 gm2 NN 1931 1939 sequence sequence NN 1939 1940 . . . 1941 1945 This this DT 1946 1954 suggests suggest VBZ 1955 1959 that that IN 1960 1963 the the DT 1964 1974 activation activation NN 1975 1984 mechanism mechanism NN 1985 1987 of of IN 1988 1991 the the DT 1992 1998 GM-CSF GM-CSF NNP 1999 2003 gene gene NN 2004 2011 through through IN 2012 2015 the the DT 2016 2026 GM2/GC-box gm2/gc-box NN 2027 2035 sequence sequence NN 2036 2038 is be VBZ 2039 2048 different different JJ 2049 2053 from from IN 2054 2058 that that DT 2059 2061 of of IN 2062 2067 genes gene NNS 2068 2076 carrying carry VBG 2077 2080 the the DT 2081 2086 kappa kappa NN 2087 2088 B B NNP 2089 2097 enhancer enhancer NN 2098 2103 alone alone RB 2103 2104 . . .