0 1 A a DT 2 7 novel novel JJ 8 12 SP-1 sp-1 NN 13 17 site site NN 18 20 in in IN 21 24 the the DT 25 30 human human JJ 31 44 interleukin-1 interleukin-1 NN 45 49 beta beta NN 50 58 promoter promoter NN 59 66 confers confer VBZ 67 79 preferential preferential JJ 80 95 transcriptional transcriptional JJ 96 104 activity activity NN 105 107 in in IN 108 121 keratinocytes keratinocyte NNS 121 122 . . . 124 126 To to TO 127 138 investigate investigate VB 139 142 the the DT 143 153 mechanisms mechanism NNS 154 156 of of IN 157 172 transcriptional transcriptional JJ 173 183 activation activation NN 184 186 of of IN 187 204 interleukin-1beta interleukin-1beta NN 205 206 ( ( ( 206 214 IL-1beta IL-1beta NNP 214 215 ) ) ) 216 218 in in IN 219 232 non-monocytic non-monocytic JJ 233 238 cells cell NNS 238 239 , , , 240 242 we we PRP 243 254 constructed construct VBD 255 256 a a DT 257 263 series series NN 264 266 of of IN 267 275 reporter reporter NN 276 284 plasmids plasmid NNS 285 289 with with IN 290 293 the the DT 294 303 bacterial bacterial JJ 304 319 chloramphenicol chloramphenicol NN 320 337 acetyltransferase acetyltransferase NN 338 342 gene gene NN 343 349 linked link VBN 350 352 to to TO 353 360 various various JJ 361 366 parts part NNS 367 369 of of IN 370 373 the the DT 374 379 human human JJ 380 388 IL-1beta IL-1beta NNP 389 397 promoter promoter NN 398 401 and and CC 402 411 performed perform VBD 412 421 transient transient JJ 422 434 transfection transfection NN 435 446 experiments experiment NNS 446 447 . . . 448 450 We we PRP 451 461 identified identify VBD 462 463 a a DT 464 472 promoter promoter NN 473 480 segment segment NN 481 485 that that WDT 486 495 activates activate VBZ 496 509 transcription transcription NN 510 514 most most RBS 515 526 efficiently efficiently RB 527 529 in in IN 530 543 keratinocytes keratinocyte NNS 543 544 . . . 545 560 Electrophoretic electrophoretic JJ 561 569 mobility mobility NN 570 575 shift shift NN 576 582 assays assay NNS 583 584 ( ( ( 584 588 EMSA EMSA NNP 588 589 ) ) ) 590 594 with with IN 595 596 a a DT 597 603 43-mer 43-mer NN 604 619 oligonucleotide oligonucleotide NN 620 627 derived derive VBN 628 632 from from IN 633 636 the the DT 637 649 functionally functionally RB 650 660 identified identify VBN 661 671 cis-acting cis-acting JJ 672 679 element element NN 680 688 revealed reveal VBD 689 697 specific specific JJ 698 707 complexes complex NNS 707 708 . . . 709 711 By by IN 712 723 competition competition NN 724 732 analysis analysis NN 733 737 with with IN 738 751 transcription transcription NN 752 758 factor factor NN 759 768 consensus consensus NN 769 777 sequence sequence NN 778 794 oligonucleotides oligonucleotide NNS 795 798 and and CC 799 801 by by IN 802 818 immunosupershift immunosupershift NN 818 819 , , , 820 833 transcription transcription NN 834 840 factor factor NN 841 845 SP-1 sp-1 NN 846 848 or or CC 849 850 a a DT 851 858 closely closely RB 859 866 related related JJ 867 874 protein protein NN 875 878 was be VBD 879 884 shown show VBN 885 887 to to TO 888 892 bind bind VB 893 895 to to TO 896 900 this this DT 901 911 regulatory regulatory JJ 912 919 element element NN 919 920 . . . 921 924 The the DT 925 932 closest close JJS 933 938 match match NN 939 941 to to TO 942 945 the the DT 946 951 known known JJ 952 956 SP-1 sp-1 NN 957 966 consensus consensus NN 967 975 sequence sequence NN 976 982 within within IN 983 986 the the DT 987 997 respective respective JJ 998 1004 region region NN 1005 1007 is be VBZ 1008 1009 a a DT 1010 1021 TCCCCTCCCCT tcccctcccct NN 1022 1027 motif motif NN 1027 1028 . . . 1029 1037 Mutation mutation NN 1038 1040 of of IN 1041 1045 this this DT 1046 1051 motif motif NN 1052 1058 almost almost RB 1059 1069 completely completely RB 1069 1070 , , , 1071 1074 and and CC 1075 1087 specifically specifically RB 1087 1088 , , , 1089 1098 abolished abolish VBD 1099 1102 the the DT 1103 1110 binding binding NN 1111 1113 of of IN 1114 1117 two two CD 1118 1130 low-mobility low-mobility JJ 1131 1140 complexes complex NNS 1141 1144 and and CC 1145 1148 led lead VBD 1149 1151 to to TO 1152 1153 a a DT 1154 1156 95 95 CD 1156 1157 % % NN 1158 1166 decrease decrease NN 1167 1169 of of IN 1170 1182 constitutive constitutive JJ 1183 1198 transcriptional transcriptional JJ 1199 1209 activation activation NN 1210 1212 of of IN 1213 1214 a a DT 1215 1223 reporter reporter NN 1224 1233 construct construct NN 1234 1242 IL-1beta IL-1beta NNP 1243 1244 ( ( ( 1244 1253 -170/+108 -170/+108 CD 1253 1254 ) ) ) 1254 1255 . . . 1256 1264 Likewise likewise RB 1264 1265 , , , 1266 1276 activation activation NN 1277 1279 of of IN 1280 1284 this this DT 1285 1293 reporter reporter NN 1294 1303 construct construct NN 1304 1306 by by IN 1307 1312 tumor tumor NN 1313 1321 necrosis necrosis NN 1322 1334 factor-alpha factor-alpha NN 1335 1343 depended depend VBD 1344 1346 on on IN 1347 1350 the the DT 1351 1355 SP-1 sp-1 NN 1356 1360 site site NN 1360 1361 . . . 1362 1367 These these DT 1368 1380 observations observation NNS 1381 1388 suggest suggest VBP 1389 1393 that that IN 1394 1395 a a DT 1396 1415 so-far-unrecognized so-far-unrecognized JJ 1416 1420 SP-1 sp-1 NN 1421 1425 site site NN 1426 1428 in in IN 1429 1432 the the DT 1433 1438 human human JJ 1439 1447 IL-1beta IL-1beta NNP 1448 1456 promoter promoter NN 1457 1460 may may MD 1461 1472 participate participate VB 1473 1475 in in IN 1476 1479 the the DT 1480 1495 transcriptional transcriptional JJ 1496 1506 regulation regulation NN 1507 1509 of of IN 1510 1514 this this DT 1515 1519 gene gene NN 1520 1522 in in IN 1523 1536 keratinocytes keratinocyte NNS 1536 1537 . . .