0 7 Binding binding NN 8 10 of of IN 11 14 YY1 yy1 NN 15 18 and and CC 19 23 Oct1 oct1 NN 24 26 to to TO 27 28 a a DT 29 34 novel novel JJ 35 42 element element NN 43 47 that that WDT 48 61 downregulates downregulate VBZ 62 72 expression expression NN 73 75 of of IN 76 80 IL-5 il-5 NN 81 83 in in IN 84 89 human human JJ 90 91 T t NN 92 97 cells cell NNS 97 98 . . . 100 110 BACKGROUND background NN 110 111 : : : 112 116 IL-5 IL-5 NNP 117 125 controls control VBZ 126 137 development development NN 138 140 of of IN 141 153 eosinophilia eosinophilia NN 154 157 and and CC 158 161 has have VBZ 162 166 been be VBN 167 172 shown show VBN 173 175 to to TO 176 178 be be VB 179 187 involved involve VBN 188 190 in in IN 191 194 the the DT 195 207 pathogenesis pathogenesis NN 208 210 of of IN 211 219 allergic allergic JJ 220 228 diseases disease NNS 228 229 . . . 230 232 In in IN 233 237 both both CC 238 244 atopic atopic JJ 245 248 and and CC 249 258 nonatopic nonatopic JJ 259 265 asthma asthma NN 265 266 , , , 267 275 elevated elevated JJ 276 280 IL-5 IL-5 NNP 281 284 has have VBZ 285 289 been be VBN 290 298 detected detect VBN 299 301 in in IN 302 312 peripheral peripheral JJ 313 318 blood blood NN 319 322 and and CC 323 326 the the DT 327 334 airways airway NNS 334 335 . . . 336 340 IL-5 IL-5 NNP 341 343 is be VBZ 344 352 produced produce VBN 353 359 mainly mainly RB 360 362 by by IN 363 372 activated activate VBN 373 374 T t NN 375 380 cells cell NNS 380 381 , , , 382 385 and and CC 386 389 its its PRP$ 390 400 expression expression NN 401 403 is be VBZ 404 413 regulated regulate VBN 414 416 at at IN 417 420 the the DT 421 436 transcriptional transcriptional JJ 437 442 level level NN 442 443 . . . 444 453 OBJECTIVE objective NN 453 454 : : : 455 459 This this DT 460 465 study study NN 466 473 focuses focus VBZ 474 476 on on IN 477 480 the the DT 481 491 functional functional JJ 492 500 analysis analysis NN 501 503 of of IN 504 507 the the DT 508 513 human human JJ 514 518 IL-5 il-5 NN 519 520 ( ( ( 520 525 hIL-5 hil-5 NN 525 526 ) ) ) 527 535 promoter promoter NN 536 539 and and CC 540 556 characterization characterization NN 557 559 of of IN 560 563 cis cis cis 564 575 -regulatory -regulatory JJ 576 584 elements element NNS 585 588 and and CC 589 602 transcription transcription NN 603 610 factors factor NNS 611 619 involved involve VBN 620 622 in in IN 623 626 the the DT 627 638 suppression suppression NN 639 641 of of IN 642 646 IL-5 IL-5 NNP 647 660 transcription transcription NN 661 663 in in IN 664 665 T t NN 666 671 cells cell NNS 671 672 . . . 673 680 METHODS methods NNS 680 681 : : : 682 689 Methods method NNS 690 694 used use VBN 695 697 in in IN 698 702 this this DT 703 708 study study NN 709 716 include include VBP 717 722 DNase dnase NN 723 724 I i CD 725 734 footprint footprint NN 735 741 assays assay NNS 741 742 , , , 743 758 electrophoretic electrophoretic JJ 759 767 mobility mobility NN 768 773 shift shift NN 774 780 assays assay NNS 780 781 , , , 782 785 and and CC 786 796 functional functional JJ 797 805 analysis analysis NN 806 808 by by IN 809 818 mammalian mammalian JJ 819 823 cell cell NN 824 836 transfection transfection NN 837 846 involving involve VBG 847 855 deletion deletion NN 856 864 analysis analysis NN 865 868 and and CC 869 882 site-directed site-directed JJ 883 894 mutagenesis mutagenesis NN 894 895 . . . 896 903 RESULTS results NNS 903 904 : : : 905 907 We we PRP 908 918 identified identify VBD 919 920 5 5 CD 921 928 protein protein NN 929 936 binding binding NN 937 944 regions region NNS 945 946 ( ( ( 946 949 BRs BR NNP 949 950 ) ) ) 951 958 located located JJ 959 965 within within IN 966 969 the the DT 970 978 proximal proximal JJ 979 984 hIL-5 hil-5 NN 985 993 promoter promoter NN 993 994 . . . 995 1005 Functional functional JJ 1006 1014 analysis analysis NN 1015 1024 indicates indicate VBZ 1025 1029 that that IN 1030 1033 the the DT 1034 1037 BRs BR NNPS 1038 1041 are be VBP 1042 1050 involved involve VBN 1051 1053 in in IN 1054 1061 control control NN 1062 1064 of of IN 1065 1070 hIL-5 hil-5 NN 1071 1079 promoter promoter NN 1080 1088 activity activity NN 1088 1089 . . . 1090 1093 Two two CD 1094 1096 of of IN 1097 1102 these these DT 1103 1110 regions region NNS 1110 1111 , , , 1112 1115 BR3 br3 NN 1116 1119 and and CC 1120 1123 BR4 br4 NN 1124 1131 located located JJ 1132 1134 at at IN 1135 1144 positions position NNS 1145 1149 -102 -102 CD 1150 1152 to to TO 1153 1156 -73 -73 CD 1156 1157 , , , 1158 1162 have have VBP 1163 1166 not not RB 1167 1177 previously previously RB 1178 1182 been be VBN 1183 1192 described describe VBN 1193 1195 as as IN 1196 1206 regulators regulator NNS 1207 1209 of of IN 1210 1214 IL-5 il-5 NN 1215 1225 expression expression NN 1226 1228 in in IN 1229 1230 T t NN 1231 1236 cells cell NNS 1236 1237 . . . 1238 1240 We we PRP 1241 1245 show show VBP 1246 1250 that that IN 1251 1254 the the DT 1255 1258 BR3 br3 NN 1259 1267 sequence sequence NN 1268 1276 contains contain VBZ 1277 1278 a a DT 1279 1284 novel novel JJ 1285 1293 negative negative JJ 1294 1304 regulatory regulatory JJ 1305 1312 element element NN 1313 1320 located located JJ 1321 1323 at at IN 1324 1333 positions position NNS 1334 1337 -90 -90 CD 1338 1340 to to TO 1341 1344 -79 -79 CD 1345 1347 of of IN 1348 1351 the the DT 1352 1357 hIL-5 hil-5 NN 1358 1366 promoter promoter NN 1366 1367 , , , 1368 1373 which which WDT 1374 1379 binds bind VBZ 1380 1384 Oct1 oct1 NN 1384 1385 , , , 1386 1398 octamer-like octamer-like JJ 1398 1399 , , , 1400 1403 and and CC 1404 1407 YY1 yy1 NN 1408 1415 nuclear nuclear JJ 1416 1423 factors factor NNS 1423 1424 . . . 1425 1437 Substitution substitution NN 1438 1447 mutations mutation NNS 1447 1448 , , , 1449 1454 which which WDT 1455 1464 abolished abolish VBD 1465 1472 binding binding NN 1473 1475 of of IN 1476 1481 these these DT 1482 1490 proteins protein NNS 1491 1493 to to TO 1494 1497 the the DT 1498 1501 BR3 br3 NN 1502 1510 sequence sequence NN 1510 1511 , , , 1512 1525 significantly significantly RB 1526 1535 increased increase VBD 1536 1541 hIL-5 hil-5 NN 1542 1550 promoter promoter NN 1551 1559 activity activity NN 1560 1562 in in IN 1563 1572 activated activate VBN 1573 1574 T t NN 1575 1580 cells cell NNS 1580 1581 . . . 1582 1592 CONCLUSION conclusion NN 1592 1593 : : : 1594 1596 We we PRP 1597 1604 suggest suggest VBP 1605 1609 that that IN 1610 1614 Oct1 Oct1 NNP 1614 1615 , , , 1616 1619 YY1 YY1 NNP 1619 1620 , , , 1621 1624 and and CC 1625 1637 octamer-like octamer-like JJ 1638 1645 factors factor NNS 1646 1653 binding bind VBG 1654 1656 to to TO 1657 1660 the the DT 1661 1668 -90/-79 -90/-79 CD 1669 1677 sequence sequence NN 1678 1684 within within IN 1685 1688 the the DT 1689 1697 proximal proximal JJ 1698 1702 IL-5 il-5 NN 1703 1711 promoter promoter NN 1712 1715 are be VBP 1716 1724 involved involve VBN 1725 1727 in in IN 1728 1739 suppression suppression NN 1740 1742 of of IN 1743 1747 IL-5 IL-5 NNP 1748 1761 transcription transcription NN 1762 1764 in in IN 1765 1766 T t NN 1767 1772 cells cell NNS 1772 1773 . . .