0 3 The the DT 4 19 immediate-early immediate-early JJ 20 24 gene gene NN 25 32 product product NN 33 38 Egr-1 egr-1 NN 39 48 regulates regulate VBZ 49 52 the the DT 53 58 human human JJ 59 72 interleukin-2 interleukin-2 NN 73 81 receptor receptor NN 82 92 beta-chain beta-chain NN 93 101 promoter promoter NN 102 109 through through IN 110 122 noncanonical noncanonical JJ 123 126 Egr Egr NNP 127 130 and and CC 131 134 Sp1 sp1 NN 135 142 binding binding NN 143 148 sites site NNS 148 149 . . . 151 154 The the DT 155 168 interleukin-2 interleukin-2 NN 169 173 IL-2 il-2 NN 174 182 receptor receptor NN 183 193 beta-chain beta-chain NN 194 195 ( ( ( 195 204 IL-2Rbeta IL-2Rbeta NNP 204 205 ) ) ) 206 208 is be VBZ 209 211 an an DT 212 221 essential essential JJ 222 231 component component NN 232 234 of of IN 235 238 the the DT 239 248 receptors receptor NNS 249 252 for for IN 253 257 IL-2 il-2 NN 258 261 and and CC 262 267 IL-15 IL-15 NNP 267 268 . . . 269 277 Although although IN 278 287 IL-2Rbeta IL-2Rbeta NNP 288 290 is be VBZ 291 305 constitutively constitutively RB 306 315 expressed express VBN 316 318 by by IN 319 330 lymphocytes lymphocyte NNS 330 331 , , , 332 335 its its PRP$ 336 346 expression expression NN 347 350 can can MD 351 353 be be VB 354 361 further further RBR 362 369 induced induce VBN 370 372 by by IN 373 374 a a DT 375 381 number number NN 382 384 of of IN 385 392 stimuli stimulus NNS 392 393 , , , 394 403 including include VBG 404 411 phorbol phorbol NN 412 424 12-myristate 12-myristate NN 425 435 13-acetate 13-acetate NN 436 437 ( ( ( 437 440 PMA PMA NNP 440 441 ) ) ) 441 442 . . . 443 445 We we PRP 446 450 have have VBP 451 454 now now RB 455 468 characterized characterize VBN 469 476 factors factor NNS 477 481 that that WDT 482 486 bind bind VBP 487 489 to to TO 490 492 an an DT 493 501 enhancer enhancer NN 502 508 region region NN 509 516 located located JJ 517 524 between between IN 525 536 nucleotides nucleotide NNS 537 541 -170 -170 CD 542 545 and and CC 546 550 -139 -139 CD 551 553 of of IN 554 557 the the DT 558 563 human human JJ 564 573 IL-2Rbeta IL-2Rbeta NNP 574 582 promoter promoter NN 582 583 . . . 584 588 Both both CC 589 592 Sp1 sp1 NN 593 596 and and CC 597 600 Sp3 sp3 NN 601 606 bound bind VBD 607 609 to to TO 610 613 the the DT 614 615 5 5 CD 615 616 ’ ' SYM 617 624 portion portion NN 625 627 of of IN 628 632 this this DT 633 639 region region NN 639 640 , , , 641 648 whereas whereas IN 649 650 a a DT 651 664 PMA-inducible pma-inducible JJ 665 671 factor factor NN 672 673 ( ( ( 673 676 PIF PIF NNP 676 677 ) ) ) 678 684 mainly mainly RB 685 690 bound bind VBD 691 693 to to TO 694 697 its its PRP$ 698 699 3 3 CD 699 700 ’ ' SYM 701 708 portion portion NN 709 712 and and CC 713 718 bound bind VBD 719 721 to to TO 722 725 the the DT 726 728 Sp Sp NNP 729 736 binding binding NN 737 743 motifs motif NNS 744 746 as as RB 747 751 well well RB 751 752 . . . 753 755 In in IN 756 762 Jurkat Jurkat NNP 763 764 T t NN 765 770 cells cell NNS 770 771 , , , 772 781 induction induction NN 782 784 of of IN 785 788 PIF PIF NNP 789 792 DNA DNA NNP 793 800 binding binding NN 801 809 activity activity NN 810 813 was be VBD 814 821 rapidly rapidly RB 822 829 induced induce VBN 829 830 , , , 831 839 required require VBN 840 842 de de FW 843 847 novo novo FW 848 855 protein protein NN 856 865 synthesis synthesis NN 865 866 , , , 867 870 and and CC 871 874 was be VBD 875 884 sustained sustained JJ 885 887 at at IN 888 889 a a DT 890 894 high high JJ 895 900 level level NN 901 904 for for IN 905 907 at at IN 908 913 least least JJS 914 916 23 23 CD 917 919 h. h. NN 920 933 Interestingly interestingly RB 933 934 , , , 935 938 PIF PIF NNP 939 942 was be VBD 943 957 constitutively constitutively RB 958 967 activated activate VBN 968 970 in in IN 971 976 human human JJ 977 983 T-cell t-cell NN 984 992 leukemia leukemia NN 993 998 virus virus NN 999 1003 type type NN 1004 1017 1-transformed 1-transformed JJ 1018 1022 MT-2 mt-2 NN 1023 1028 cells cell NNS 1028 1029 . . . 1030 1032 In in IN 1033 1037 this this DT 1038 1043 paper paper NN 1043 1044 , , , 1045 1047 we we PRP 1048 1059 demonstrate demonstrate VBP 1060 1064 that that IN 1065 1068 PIF PIF NNP 1069 1071 is be VBZ 1072 1077 Egr-1 egr-1 NN 1078 1083 based base VBN 1084 1086 on on IN 1087 1090 its its PRP$ 1091 1102 recognition recognition NN 1103 1105 by by IN 1106 1116 anti-Egr-1 anti-egr-1 JJ 1117 1125 antisera antiserum NNS 1126 1128 in in IN 1129 1132 gel gel NN 1133 1141 mobility mobility NN 1142 1147 shift shift NN 1148 1154 assays assay NNS 1154 1155 , , , 1156 1160 even even RB 1161 1167 though though IN 1168 1171 the the DT 1172 1181 IL-2Rbeta IL-2Rbeta NNP 1182 1185 DNA DNA NNP 1186 1193 binding binding NN 1194 1199 motif motif NN 1200 1208 differed differ VBD 1209 1222 substantially substantially RB 1223 1227 from from IN 1228 1231 the the DT 1232 1241 canonical canonical JJ 1242 1247 Egr-1 egr-1 NN 1248 1255 binding binding NN 1256 1260 site site NN 1260 1261 . . . 1262 1264 In in IN 1265 1273 addition addition NN 1273 1274 , , , 1275 1280 Egr-1 egr-1 NN 1281 1286 bound bind VBD 1287 1289 to to TO 1290 1293 the the DT 1294 1296 Sp Sp NNP 1297 1304 binding binding NN 1305 1309 site site NN 1309 1310 . . . 1311 1313 In in IN 1314 1320 Jurkat Jurkat NNP 1321 1326 cells cell NNS 1326 1327 , , , 1328 1332 both both DT 1333 1338 sites site NNS 1339 1343 were be VBD 1344 1352 required require VBN 1353 1356 for for IN 1357 1364 maximal maximal JJ 1365 1374 IL-2Rbeta IL-2Rbeta NNP 1375 1383 promoter promoter NN 1384 1392 activity activity NN 1392 1393 , , , 1394 1397 and and CC 1398 1400 in in IN 1401 1407 HeLaS3 helas3 NN 1408 1413 cells cell NNS 1413 1414 , , , 1415 1427 transfection transfection NN 1428 1430 of of IN 1431 1436 Egr-1 Egr-1 NNP 1437 1442 could could MD 1443 1448 drive drive VB 1449 1457 activity activity NN 1458 1460 of of IN 1461 1462 a a DT 1463 1471 reporter reporter NN 1472 1481 construct construct NN 1482 1492 containing contain VBG 1493 1497 both both DT 1498 1503 sites site NNS 1503 1504 . . . 1505 1513 Moreover moreover RB 1513 1514 , , , 1515 1518 Sp1 Sp1 NNP 1519 1522 and and CC 1523 1528 Egr-1 Egr-1 NNP 1529 1534 could could MD 1535 1539 form form VB 1540 1541 a a DT 1542 1549 complex complex NN 1550 1554 with with IN 1555 1563 kinetics kinetic NNS 1564 1568 that that WDT 1569 1579 correlated correlate VBD 1580 1584 with with IN 1585 1588 the the DT 1589 1599 production production NN 1600 1602 of of IN 1603 1608 Egr-1 egr-1 NN 1609 1611 in in IN 1612 1618 Jurkat Jurkat NNP 1619 1624 cells cell NNS 1625 1629 upon upon IN 1630 1633 PMA PMA NNP 1634 1645 stimulation stimulation NN 1645 1646 . . . 1647 1651 Thus thus RB 1651 1652 , , , 1653 1656 Sp1 Sp1 NNP 1657 1660 and and CC 1661 1666 Egr-1 egr-1 NN 1667 1677 physically physically RB 1678 1681 and and CC 1682 1694 functionally functionally RB 1695 1704 cooperate cooperate VBP 1705 1707 to to TO 1708 1715 mediate mediate VB 1716 1723 maximal maximal JJ 1724 1733 IL-2Rbeta IL-2Rbeta NNP 1734 1742 promoter promoter NN 1743 1751 activity activity NN 1751 1752 . . .