0 12 Dimerization dimerization NN 13 15 of of IN 16 22 NF-KB2 nf-kb2 NN 23 27 with with IN 28 37 RelA(p65) RelA(p65) NNP 38 47 regulates regulate VBZ 48 51 DNA dna NN 52 59 binding binding NN 59 60 , , , 61 76 transcriptional transcriptional JJ 77 87 activation activation NN 87 88 , , , 89 92 and and CC 93 103 inhibition inhibition NN 104 106 by by IN 107 109 an an DT 110 111 I I NNP 112 117 kappa kappa NN 118 125 B-alpha B-alpha NNP 126 127 ( ( ( 127 132 MAD-3 MAD-3 NNP 132 133 ) ) ) 133 134 . . . 136 145 Inducible inducible JJ 146 156 expression expression NN 157 159 of of IN 160 165 human human JJ 166 182 immunodeficiency immunodeficiency NN 183 188 virus virus NN 189 190 ( ( ( 190 193 HIV HIV NNP 193 194 ) ) ) 195 197 is be VBZ 198 207 regulated regulate VBN 208 210 by by IN 211 212 a a DT 213 221 cellular cellular JJ 222 235 transcription transcription NN 236 242 factor factor NN 242 243 , , , 244 251 nuclear nuclear JJ 252 258 factor factor NN 259 264 kappa kappa NN 265 266 B B NNP 267 268 ( ( ( 268 276 NF-kappa NF-kappa NNP 277 278 B B NNP 278 279 ) ) ) 279 280 . . . 281 289 NF-kappa NF-kappa NNP 290 291 B B NNP 292 294 is be VBZ 295 303 composed compose VBN 304 306 of of IN 307 315 distinct distinct JJ 316 324 subunits subunit NNS 324 325 ; ; : 326 330 five five CD 331 342 independent independent JJ 343 348 genes gene NNS 348 349 , , , 350 361 NFKB1(p105) NFKB1(p105) NNP 361 362 , , , 363 374 NFKB2(p100) NFKB2(p100) NNP 374 375 , , , 376 385 RelA(p65) RelA(p65) NNP 385 386 , , , 387 392 c-rel c-rel NN 393 396 and and CC 397 401 relB relB NNP 401 402 , , , 403 407 that that WDT 408 414 encode encode VBP 415 422 related related JJ 423 431 proteins protein NNS 432 436 that that WDT 437 441 bind bind VBP 442 444 to to TO 445 450 kappa kappa NN 451 452 B B NNP 453 456 DNA DNA NNP 457 465 elements element NNS 466 470 have have VBP 471 475 been be VBN 476 484 isolated isolate VBN 484 485 . . . 486 488 We we PRP 489 493 have have VBP 494 504 previously previously RB 505 510 found find VBN 511 515 that that IN 516 530 NFKB2(p49/p52) nfkb2(p49/p52) NN 531 535 acts act VBZ 536 538 in in IN 539 546 concert concert NN 547 551 with with IN 552 561 RelA(p65) RelA(p65) NNP 562 564 to to TO 565 574 stimulate stimulate VB 575 578 the the DT 579 582 HIV HIV NNP 583 591 enhancer enhancer NN 592 594 in in IN 595 601 Jurkat Jurkat NNP 602 612 T-leukemia T-leukemia NNP 613 618 cells cell NNS 618 619 . . . 620 624 Here here RB 625 627 we we PRP 628 635 examine examine VBP 636 639 the the DT 640 651 biochemical biochemical JJ 652 657 basis basis NN 658 661 for for IN 662 665 the the DT 666 681 transcriptional transcriptional JJ 682 692 regulation regulation NN 693 695 of of IN 696 699 HIV HIV NNP 700 702 by by IN 703 708 NFKB2 nfkb2 NN 708 709 . . . 710 715 Using use VBG 716 725 Scatchard Scatchard NNP 726 734 analysis analysis NN 734 735 , , , 736 738 we we PRP 739 743 have have VBP 744 754 determined determine VBN 755 758 the the DT 759 771 dissociation dissociation NN 772 781 constants constant NNS 782 784 of of IN 785 796 homodimeric homodimeric JJ 797 800 p49 p49 NN 801 804 and and CC 805 818 heterodimeric heterodimeric JJ 819 826 p49/p65 p49/p65 NN 827 830 for for IN 831 838 binding bind VBG 839 841 to to TO 842 845 the the DT 846 849 HIV HIV NNP 850 855 kappa kappa NN 856 857 B B NNP 858 863 site. site. NN 864 867 p49 p49 NN 868 871 has have VBZ 872 873 a a DT 874 887 approximately approximately RB 888 901 18-fold-lower 18-fold-lower JJ 902 910 affinity affinity NN 911 914 for for IN 915 918 the the DT 919 922 HIV HIV NNP 923 928 kappa kappa NN 929 930 B B NNP 931 935 site site NN 936 937 ( ( ( 937 939 KD kd NN 940 941 = = SYM 942 946 69.1 69.1 CD 947 949 pM pm NN 949 950 ) ) ) 951 955 than than IN 956 960 does do VBZ 961 964 the the DT 965 978 approximately approximately RB 979 985 50-kDa 50-kda JJ 986 993 protein protein NN 994 1004 NFKB1(p50) nfkb1(p50) NN 1005 1012 derived derive VBN 1013 1017 from from IN 1018 1022 p105 p105 NN 1023 1024 ( ( ( 1024 1026 KD kd NN 1027 1028 = = SYM 1029 1032 3.9 3.9 CD 1033 1035 pM pM NNP 1035 1036 ) ) ) 1036 1037 . . . 1038 1040 In in IN 1041 1049 contrast contrast NN 1049 1050 , , , 1051 1054 the the DT 1055 1063 affinity affinity NN 1064 1066 of of IN 1067 1080 heterodimeric heterodimeric JJ 1081 1101 NFKB2(p49)/RelA(p65) nfkb2(p49)/rela(p65) NN 1102 1105 for for IN 1106 1110 this this DT 1111 1115 site site NN 1116 1118 is be VBZ 1119 1132 approximately approximately RB 1133 1139 6-fold 6-fold RB 1140 1146 higher high JJR 1147 1148 ( ( ( 1148 1150 KD kd NN 1151 1152 = = SYM 1153 1157 11.8 11.8 CD 1158 1160 pM pm NN 1160 1161 ) ) ) 1162 1166 than than IN 1167 1171 that that DT 1172 1174 of of IN 1175 1178 p49 p49 NN 1179 1184 alone alone RB 1184 1185 . . . 1186 1196 Consistent consistent JJ 1197 1201 with with IN 1202 1207 these these DT 1208 1216 findings finding NNS 1216 1217 , , , 1218 1220 in in FW 1221 1226 vitro vitro FW 1227 1240 transcription transcription NN 1241 1244 was be VBD 1245 1255 stimulated stimulate VBN 1256 1263 18-fold 18-fold RB 1264 1266 by by IN 1267 1270 the the DT 1271 1279 addition addition NN 1280 1282 of of IN 1283 1292 preformed preformed JJ 1292 1293 , , , 1294 1307 heterodimeric heterodimeric JJ 1308 1328 NFKB2(p49)/RelA(p65) nfkb2(p49)/rela(p65) NN 1329 1336 protein protein NN 1336 1337 . . . 1338 1353 Transcriptional transcriptional JJ 1354 1364 activation activation NN 1365 1367 of of IN 1368 1371 the the DT 1372 1375 HIV HIV NNP 1376 1384 enhancer enhancer NN 1385 1388 was be VBD 1389 1393 also also RB 1394 1401 subject subject JJ 1402 1404 to to TO 1405 1415 regulation regulation NN 1416 1418 by by IN 1419 1427 recently recently RB 1428 1434 cloned clone VBN 1435 1436 I i NN 1437 1442 kappa kappa NN 1443 1457 B-alpha(MAD-3) b-alpha(mad-3) NN 1457 1458 . . . 1459 1470 Recombinant recombinant JJ 1471 1472 I i NN 1473 1478 kappa kappa NN 1479 1493 B-alpha(MAD-3) b-alpha(mad-3) NN 1494 1503 inhibited inhibit VBD 1504 1507 the the DT 1508 1511 DNA dna NN 1512 1519 binding binding NN 1520 1528 activity activity NN 1529 1531 of of IN 1532 1535 p65 p65 NN 1535 1536 , , , 1537 1544 p49/p65 p49/p65 NN 1544 1545 , , , 1546 1549 and and CC 1550 1557 p50/p65 p50/p65 NN 1558 1561 but but CC 1562 1572 stimulated stimulate VBD 1573 1576 the the DT 1577 1584 binding binding NN 1585 1587 of of IN 1588 1598 NFKB2(p49) NFKB2(p49) NNP 1599 1601 or or CC 1602 1612 NFKB1(p50) NFKB1(p50) NNP 1612 1613 . . . 1614 1624 Functional functional JJ 1625 1635 activation activation NN 1636 1638 of of IN 1639 1641 an an DT 1642 1645 HIV HIV NNP 1646 1654 reporter reporter NN 1655 1662 plasmid plasmid NN 1663 1665 by by IN 1666 1673 p49/p65 p49/p65 NN 1674 1676 in in IN 1677 1688 transiently transiently RB 1689 1700 transfected transfecte VBN 1701 1707 Jurkat Jurkat NNP 1708 1718 T-leukemia T-leukemia NNP 1719 1724 cells cell NNS 1725 1728 was be VBD 1729 1733 also also RB 1734 1743 inhibited inhibit VBN 1744 1746 by by IN 1747 1759 coexpression coexpression NN 1760 1762 of of IN 1763 1768 MAD-3 MAD-3 NNP 1768 1769 . . . 1770 1771 ( ( ( 1771 1779 ABSTRACT abstract NN 1780 1789 TRUNCATED truncated VBN 1790 1792 AT at IN 1793 1796 250 250 CD 1797 1802 WORDS words NNS 1802 1803 ) ) )