0 8 Multiple multiple JJ 9 17 proteins protein NNS 18 26 interact interact VBP 27 31 with with IN 32 35 the the DT 36 43 nuclear nuclear JJ 44 54 inhibitory inhibitory JJ 55 62 protein protein NN 63 72 repressor repressor NN 73 80 element element NN 81 83 in in IN 84 87 the the DT 88 93 human human JJ 94 107 interleukin-3 interleukin-3 NN 108 116 promoter promoter NN 116 117 . . . 119 120 T t NN 121 125 cell cell NN 126 136 expression expression NN 137 139 of of IN 140 151 interleukin interleukin NN 152 153 3 3 CD 154 155 ( ( ( 155 159 IL-3 il-3 NN 159 160 ) ) ) 161 163 is be VBZ 164 172 directed direct VBN 173 175 by by IN 176 184 positive positive JJ 185 188 and and CC 189 197 negative negative JJ 198 208 cis-acting cis-acting JJ 209 212 DNA dna NN 213 221 elements element NNS 222 231 clustered cluster VBN 232 238 within within IN 239 242 300 300 CD 243 247 base base NN 248 253 pairs pair NNS 254 256 of of IN 257 260 the the DT 261 276 transcriptional transcriptional JJ 277 282 start start NN 283 287 site site NN 287 288 . . . 289 290 A a DT 291 297 strong strong JJ 298 307 repressor repressor NN 308 315 element element NN 315 316 , , , 317 323 termed term VBN 324 331 nuclear nuclear JJ 332 342 inhibitory inhibitory JJ 343 350 protein protein NN 351 352 ( ( ( 352 355 NIP NIP NNP 355 356 ) ) ) 356 357 , , , 358 361 was be VBD 362 372 previously previously RB 373 379 mapped map VBN 380 382 to to TO 383 384 a a DT 385 392 segment segment NN 393 395 of of IN 396 399 the the DT 400 404 IL-3 il-3 NN 405 413 promoter promoter NN 414 421 between between IN 422 433 nucleotides nucleotide NNS 434 438 -271 -271 CD 439 442 and and CC 443 447 -250 -250 CD 447 448 . . . 449 459 Functional functional JJ 460 476 characterization characterization NN 477 479 of of IN 480 484 this this DT 485 492 element element NN 493 505 demonstrates demonstrate VBZ 506 510 that that IN 511 513 it it PRP 514 517 can can MD 518 525 mediate mediate VB 526 536 repression repression NN 537 541 when when WRB 542 548 linked link VBN 549 551 in in IN 552 555 cis cis NN 556 558 to to TO 559 560 a a DT 561 573 heterologous heterologous JJ 574 582 promoter promoter NN 582 583 . . . 584 587 DNA DNA NNP 588 595 binding binding NN 596 607 experiments experiment NNS 608 612 were be VBD 613 620 carried carry VBN 621 624 out out RP 625 627 to to TO 628 640 characterize characterize VB 641 644 the the DT 645 654 repressor repressor NN 655 663 activity activity NN 663 664 . . . 665 670 Using use VBG 671 678 varying vary VBG 679 689 conditions condition NNS 689 690 , , , 691 696 three three CD 697 705 distinct distinct JJ 706 715 complexes complex NNS 716 720 were be VBD 721 726 shown show VBN 727 729 to to TO 730 738 interact interact VB 739 751 specifically specifically RB 752 756 with with IN 757 760 the the DT 761 764 NIP NIP NNP 765 771 region region NN 771 772 , , , 773 781 although although IN 782 786 only only RB 787 790 one one CD 791 801 correlates correlate VBZ 802 806 with with IN 807 816 repressor repressor NN 817 825 activity activity NN 825 826 . . . 827 834 Complex Complex NNP 835 836 1 1 CD 837 844 results result NNS 845 849 from from IN 850 857 binding binding NN 858 860 of of IN 861 862 a a DT 863 873 ubiquitous ubiquitous JJ 874 885 polypeptide polypeptide NN 886 890 that that WDT 891 901 recognizes recognize VBZ 902 905 the the DT 906 907 3 3 CD 907 908 ’ ' SYM 909 916 portion portion NN 917 919 of of IN 920 924 this this DT 925 933 sequence sequence NN 934 937 and and CC 938 940 is be VBZ 941 944 not not RB 945 953 required require VBN 954 957 for for IN 958 968 repression repression NN 968 969 . . . 970 977 Complex complex NN 978 979 2 2 CD 980 991 corresponds correspond VBZ 992 994 to to TO 995 1002 binding binding NN 1003 1005 of of IN 1006 1019 transcription transcription NN 1020 1026 factor factor NN 1027 1028 ( ( ( 1028 1036 upstream upstream JJ 1037 1048 stimulatory stimulatory JJ 1049 1055 factor factor NN 1055 1056 ) ) ) 1057 1059 to to TO 1060 1062 an an DT 1063 1068 E-box e-box NN 1069 1074 motif motif NN 1075 1077 in in IN 1078 1081 the the DT 1082 1083 5 5 CD 1083 1084 ’ ' SYM 1085 1092 portion portion NN 1093 1095 of of IN 1096 1099 the the DT 1100 1103 NIP NIP NNP 1104 1110 region region NN 1110 1111 . . . 1112 1115 DNA DNA NNP 1116 1123 binding binding NN 1124 1135 specificity specificity NN 1136 1138 of of IN 1139 1146 complex complex NN 1147 1148 3 3 CD 1149 1157 overlaps overlap NNS 1158 1162 with with IN 1163 1167 that that DT 1168 1170 of of IN 1171 1179 upstream upstream JJ 1180 1191 stimulatory stimulatory JJ 1192 1198 factor factor NN 1199 1202 but but CC 1203 1205 is be VBZ 1206 1213 clearly clearly RB 1214 1222 distinct distinct JJ 1222 1223 . . . 1224 1226 To to TO 1227 1236 determine determine VB 1237 1242 which which WDT 1243 1245 of of IN 1246 1249 the the DT 1250 1256 latter latter JJ 1257 1260 two two CD 1261 1270 complexes complex NNS 1271 1281 represents represent VBZ 1282 1285 NIP NIP NNP 1286 1294 activity activity NN 1294 1295 , , , 1296 1298 we we PRP 1299 1311 incorporated incorporate VBD 1312 1317 small small JJ 1318 1329 alterations alteration NNS 1330 1334 into into IN 1335 1338 the the DT 1339 1342 NIP NIP NNP 1343 1347 site site NN 1348 1350 of of IN 1351 1353 an an DT 1354 1358 IL-3 il-3 NN 1359 1374 promoter-linked promoter-linked JJ 1375 1383 reporter reporter NN 1384 1393 construct construct NN 1394 1397 and and CC 1398 1406 examined examine VBN 1407 1412 their their PRP$ 1413 1420 effects effect NNS 1421 1423 on on IN 1424 1436 NIP-mediated nip-mediated JJ 1437 1447 repression repression NN 1447 1448 . . . 1449 1459 Functional functional JJ 1460 1471 specificity specificity NN 1472 1475 for for IN 1476 1486 repression repression NN 1487 1494 matches match VBZ 1495 1498 the the DT 1499 1502 DNA dna NN 1503 1510 binding binding NN 1511 1522 specificity specificity NN 1523 1525 of of IN 1526 1533 complex complex NN 1534 1535 3 3 CD 1535 1536 ; ; : 1537 1541 both both CC 1542 1551 repressor repressor NN 1552 1560 activity activity NN 1561 1564 and and CC 1565 1572 complex complex NN 1573 1574 3 3 CD 1575 1582 binding binding NN 1583 1590 require require VBP 1591 1594 the the DT 1595 1604 consensus consensus NN 1605 1613 sequence sequence NN 1614 1623 CTCACNTNC CTCACNTNC NNP 1623 1624 . . .