0 11 Involvement involvement NN 12 14 of of IN 15 24 NF-kappaB NF-kappaB NNP 25 32 p50/p65 p50/p65 NN 33 44 heterodimer heterodimer NN 45 47 in in IN 48 58 activation activation NN 59 61 of of IN 62 65 the the DT 66 71 human human JJ 72 93 pro-interleukin-1beta pro-interleukin-1beta NN 94 98 gene gene NN 99 101 at at IN 102 105 two two CD 106 116 subregions subregion NNS 117 119 of of IN 120 123 the the DT 124 132 upstream upstream JJ 133 141 enhancer enhancer NN 142 149 element element NN 149 150 . . . 152 153 A a DT 154 160 region region NN 161 173 between-3134 between-3134 CD 174 177 and and CC 178 183 -2729 -2729 CD 184 186 bp bp NN 187 195 upstream upstream RB 196 200 from from IN 201 204 the the DT 205 218 transcription transcription NN 219 223 site site NN 224 226 of of IN 227 230 the the DT 231 236 human human JJ 237 252 pro-interleukin pro-interleukin JJ 253 258 1beta 1beta NN 259 260 ( ( ( 260 271 proIL-1beta proil-1beta NN 271 272 ) ) ) 273 277 gene gene NN 278 281 was be VBD 282 292 identified identify VBN 293 295 as as IN 296 298 an an DT 299 313 LPS-responsive lps-responsive JJ 314 322 enhancer enhancer NN 323 330 element element NN 330 331 . . . 332 334 In in IN 335 339 this this DT 340 345 study study NN 345 346 , , , 347 350 the the DT 351 360 influence influence NN 361 363 of of IN 364 367 the the DT 368 377 sequences sequence NNS 378 385 located located JJ 386 393 between between IN 394 399 -3134 -3134 CD 400 403 and and CC 404 409 -2987 -2987 CD 410 412 on on IN 413 416 the the DT 417 432 transcriptional transcriptional JJ 433 441 activity activity NN 442 444 of of IN 445 448 the the DT 449 460 proIL-1beta proil-1beta NN 461 465 gene gene NN 466 468 in in IN 469 483 LPS-stimulated lps-stimulated JJ 484 487 Raw Raw NNP 488 493 264.7 264.7 CD 494 499 cells cell NNS 500 503 was be VBD 504 512 examined examine VBN 513 515 in in IN 516 522 detail detail NN 522 523 . . . 524 527 The the DT 528 535 results result NNS 536 544 obtained obtain VBN 545 547 by by IN 548 557 transient transient JJ 558 570 transfection transfection NN 571 573 of of IN 574 577 fos fos NN 578 582 -CAT -CAT NNP 583 593 constructs construct NNS 594 598 that that WDT 599 608 contained contain VBD 609 615 serial serial JJ 616 627 5’-deletion 5'-deletion JJ 628 637 mutations mutation NNS 638 644 showed show VBD 645 649 that that IN 650 653 the the DT 654 660 region region NN 661 668 between between IN 669 674 -3134 -3134 CD 675 678 and and CC 679 684 -3059 -3059 CD 685 692 appears appear VBZ 693 695 to to TO 696 698 be be VB 699 707 required require VBN 708 711 for for IN 712 715 the the DT 716 725 induction induction NN 726 728 of of IN 729 742 transcription transcription NN 743 745 by by IN 746 749 LPS LPS NNP 749 750 . . . 751 754 Gel Gel NNP 755 760 shift shift NN 761 766 assay assay NN 767 774 studies study NNS 775 779 with with IN 780 789 synthetic synthetic JJ 790 806 oligonucleotides oligonucleotide NNS 807 820 corresponding correspond VBG 821 823 to to TO 824 831 partial partial JJ 832 841 sequences sequence NNS 842 844 of of IN 845 848 the the DT 849 855 latter latter JJ 856 862 region region NN 863 866 and and CC 867 874 nuclear nuclear JJ 875 883 extracts extract NNS 884 888 from from IN 889 899 stimulated stimulate VBN 900 905 cells cell NNS 906 914 revealed reveal VBD 915 923 specific specific JJ 924 931 protein protein NN 932 939 binding binding NN 940 945 sites site NNS 946 953 between between IN 954 959 -3110 -3110 CD 960 963 and and CC 964 969 -3090 -3090 CD 970 973 and and CC 974 981 between between IN 982 987 -3079 -3079 CD 988 991 and and CC 992 997 -3059 -3059 CD 997 998 . . . 999 1004 These these DT 1005 1013 specific specific JJ 1014 1022 bindings binding NNS 1023 1027 were be VBD 1028 1032 time time NN 1033 1036 and and CC 1037 1040 LPS LPS NNP 1041 1045 dose dose NN 1046 1055 dependent dependent JJ 1055 1056 . . . 1057 1060 The the DT 1061 1068 results result NNS 1069 1071 of of IN 1072 1082 supershift supershift NN 1083 1091 analysis analysis NN 1092 1097 using use VBG 1098 1106 specific specific JJ 1107 1117 antibodies antibody NNS 1118 1125 against against IN 1126 1139 transcription transcription NN 1140 1147 factors factor NNS 1148 1157 suggested suggest VBD 1158 1162 that that IN 1163 1167 both both CC 1168 1175 binding binding NN 1176 1185 complexes complex NNS 1186 1195 contained contain VBD 1196 1199 the the DT 1200 1209 NF-kappaB NF-kappaB NNP 1210 1220 components component NNS 1221 1224 p50 p50 NN 1225 1228 and and CC 1229 1232 p65 p65 NN 1232 1233 , , , 1234 1237 and and CC 1238 1241 did do VBD 1242 1245 not not RB 1246 1253 contain contain VB 1254 1259 other other JJ 1260 1269 NF-kappaB NF-kappaB NNP 1270 1278 proteins protein NNS 1279 1280 ( ( ( 1280 1283 p52 p52 NN 1283 1284 , , , 1285 1290 c-Rel c-rel NN 1290 1291 , , , 1292 1295 Rel Rel NNP 1296 1297 B B NNP 1297 1298 ) ) ) 1298 1299 , , , 1300 1304 AP-1 ap-1 NN 1305 1313 proteins protein NNS 1314 1315 ( ( ( 1315 1320 c-Fos c-fos NN 1320 1321 , , , 1322 1327 C-Jun C-Jun NNP 1327 1328 ) ) ) 1328 1329 , , , 1330 1334 CREB CREB NNP 1335 1337 or or CC 1338 1347 C/EBPbeta C/EBPbeta NNP 1348 1349 ( ( ( 1349 1355 NF-IL6 NF-IL6 NNP 1355 1356 ) ) ) 1356 1357 . . . 1358 1366 Mutation mutation NN 1367 1369 of of IN 1370 1376 either either DT 1377 1379 of of IN 1380 1383 the the DT 1384 1392 putative putative JJ 1393 1410 NF-kappaB-binding nf-kappab-binding JJ 1411 1416 sites site NNS 1417 1419 in in IN 1420 1423 the the DT 1424 1432 enhancer enhancer NN 1433 1440 element element NN 1441 1450 decreased decrease VBD 1451 1454 the the DT 1455 1469 LPS-stimulated lps-stimulated JJ 1470 1485 transcriptional transcriptional JJ 1486 1494 activity activity NN 1494 1495 . . . 1496 1501 These these DT 1502 1506 data datum NNS 1507 1516 indicated indicate VBD 1517 1521 that that IN 1522 1525 two two CD 1526 1543 NF-kappaB-binding nf-kappab-binding JJ 1544 1549 sites site NNS 1549 1550 , , , 1551 1556 which which WDT 1557 1560 are be VBP 1561 1568 located located JJ 1569 1576 between between IN 1577 1582 -3134 -3134 CD 1583 1586 and and CC 1587 1592 -3059 -3059 CD 1592 1593 , , , 1594 1597 are be VBP 1598 1606 critical critical JJ 1607 1610 for for IN 1611 1614 the the DT 1615 1625 activation activation NN 1626 1628 of of IN 1629 1640 proIL-1beta proil-1beta NN 1641 1645 gene gene NN 1646 1659 transcription transcription NN 1659 1660 . . . 1661 1670 Copyright Copyright NNP 1671 1675 1999 1999 CD 1676 1684 Academic Academic NNP 1685 1690 Press Press NNP 1690 1691 . . .