0 6 Relief Relief NNP 7 9 of of IN 10 16 cyclin cyclin NN 17 18 A a NN 19 23 gene gene NN 24 39 transcriptional transcriptional JJ 40 50 inhibition inhibition NN 51 57 during during IN 58 68 activation activation NN 69 71 of of IN 72 77 human human JJ 78 85 primary primary JJ 86 87 T t NN 88 99 lymphocytes lymphocyte NNS 100 103 via via IN 104 107 CD2 cd2 NN 108 111 and and CC 112 116 CD28 cd28 NN 117 125 adhesion adhesion NN 126 135 molecules molecule NNS 135 136 . . . 138 144 Cyclin Cyclin NNP 145 146 A a NN 147 160 transcription transcription NN 161 163 is be VBZ 164 168 cell cell NN 169 174 cycle cycle NN 175 184 regulated regulate VBN 185 188 and and CC 189 196 induced induce VBN 197 199 by by IN 200 204 cell cell NN 205 218 proliferative proliferative JJ 219 226 signals signal NNS 226 227 . . . 228 230 To to TO 231 241 understand understand VB 242 245 the the DT 246 256 mechanisms mechanism NNS 257 267 underlined underline VBN 268 270 in in IN 271 275 this this DT 276 286 regulation regulation NN 287 289 in in IN 290 296 normal normal JJ 297 302 human human JJ 303 308 cells cell NNS 308 309 , , , 310 312 we we PRP 313 317 have have VBP 318 326 analysed analyse VBN 327 329 in in FW 330 334 vivo vivo FW 335 346 protein-DNA protein-dna JJ 347 359 interactions interaction NNS 360 362 at at IN 363 366 the the DT 367 373 Cyclin Cyclin NNP 374 375 A a NN 376 381 locus locus NN 382 384 in in IN 385 392 primary primary JJ 393 394 T t NN 395 406 lymphocytes lymphocyte NNS 406 407 . . . 408 419 Stimulation stimulation NN 420 422 of of IN 423 431 purified purify VBN 432 433 T t NN 434 445 lymphocytes lymphocyte NNS 446 448 by by IN 449 450 a a DT 451 462 combination combination NN 463 465 of of IN 466 476 monoclonal monoclonal JJ 477 487 antibodies antibody NNS 488 496 directed direct VBN 497 499 at at IN 500 503 CD2 cd2 NN 504 507 and and CC 508 512 CD28 cd28 NN 513 521 adhesion adhesion NN 522 531 molecules molecule NNS 532 537 gives give VBZ 538 542 rise rise NN 543 545 to to TO 546 547 a a DT 548 552 long long JJ 553 560 lasting lasting JJ 561 574 proliferation proliferation NN 575 577 in in IN 578 581 the the DT 582 589 absence absence NN 590 592 of of IN 593 602 accessory accessory NN 603 608 cells cell NNS 608 609 . . . 610 616 Cyclin Cyclin NNP 617 618 A A NNP 619 622 was be VBD 623 631 observed observe VBN 632 637 after after IN 638 639 4 4 CD 640 644 days day NNS 645 647 of of IN 648 661 costimulation costimulation NN 662 666 with with IN 667 671 anti anti JJ 672 675 CD2 cd2 NN 676 677 + + CC 678 682 CD28 cd28 NN 683 690 whereas whereas IN 691 702 stimulation stimulation NN 703 705 by by IN 706 710 anti anti JJ 711 714 CD2 cd2 NN 715 717 or or CC 718 722 anti anti JJ 723 727 CD28 cd28 NN 728 733 alone alone RB 734 737 was be VBD 738 741 not not RB 742 751 effective effective JJ 751 752 . . . 753 755 In in FW 756 760 vivo vivo FW 761 768 genomic genomic JJ 769 772 DMS DMS NNP 773 785 footprinting footprinting NN 786 794 revealed reveal VBD 795 803 upstream upstream RB 804 806 of of IN 807 810 the the DT 811 816 major major JJ 817 830 transcription transcription NN 831 841 initiation initiation NN 842 847 sites site NNS 847 848 , , , 849 852 the the DT 853 861 presence presence NN 862 864 of of IN 865 867 at at IN 868 873 least least JJS 874 879 three three CD 880 887 protein protein NN 888 895 binding binding NN 896 901 sites site NNS 901 902 , , , 903 906 two two CD 907 909 of of IN 910 915 which which WDT 916 920 were be VBD 921 935 constitutively constitutively RB 936 944 occupied occupy VBN 944 945 . . . 946 950 They they PRP 951 955 bind bind VBP 956 958 in in FW 959 964 vitro vitro FW 965 977 respectively respectively RB 978 983 ATF-1 atf-1 NN 984 987 and and CC 988 992 NF-Y nf-y NN 993 1001 proteins protein NNS 1001 1002 . . . 1003 1006 The the DT 1007 1012 third third JJ 1013 1017 site site NN 1018 1021 was be VBD 1022 1030 occupied occupy VBN 1031 1033 in in IN 1034 1043 quiescent quiescent JJ 1044 1049 cells cell NNS 1050 1052 or or CC 1053 1055 in in IN 1056 1061 cells cell NNS 1062 1072 stimulated stimulate VBN 1073 1075 by by IN 1076 1080 anti anti JJ 1081 1084 CD2 cd2 NN 1085 1087 or or CC 1088 1092 anti anti JJ 1093 1097 CD28 cd28 NN 1098 1103 alone alone RB 1103 1104 . . . 1105 1108 The the DT 1109 1118 mitogenic mitogenic JJ 1119 1130 combination combination NN 1131 1133 of of IN 1134 1138 anti anti JJ 1139 1142 CD2 cd2 NN 1143 1144 + + CC 1145 1149 anti anti JJ 1150 1154 CD28 cd28 NN 1155 1163 released release VBD 1164 1167 the the DT 1168 1177 footprint footprint NN 1178 1180 as as IN 1181 1186 cells cell NNS 1187 1191 were be VBD 1192 1201 committed commit VBN 1202 1204 to to TO 1205 1218 proliferation proliferation NN 1218 1219 . . . 1220 1230 Consistent consistent JJ 1231 1235 with with IN 1236 1242 theses theses DT 1243 1250 results result NNS 1250 1251 , , , 1252 1259 nuclear nuclear JJ 1260 1268 extracts extract NNS 1269 1277 prepared prepare VBN 1278 1282 from from IN 1283 1292 quiescent quiescent JJ 1293 1298 cells cell NNS 1299 1305 formed form VBD 1306 1307 a a DT 1308 1316 specific specific JJ 1317 1324 complex complex NN 1325 1329 with with IN 1330 1334 this this DT 1335 1342 element element NN 1342 1343 , , , 1344 1351 whereas whereas IN 1352 1360 extracts extract NNS 1361 1369 prepared prepare VBN 1370 1374 from from IN 1375 1380 cells cell NNS 1381 1388 treated treat VBN 1389 1393 with with IN 1394 1398 anti anti JJ 1399 1402 CD2 cd2 NN 1403 1404 + + CC 1405 1409 anti anti JJ 1410 1414 CD28 cd28 NN 1415 1421 failed fail VBD 1422 1424 to to TO 1425 1427 do do VB 1428 1430 so so RB 1431 1436 after after IN 1437 1442 cells cell NNS 1443 1450 entered enter VBD 1451 1452 a a DT 1453 1466 proliferative proliferative JJ 1467 1472 state state NN 1472 1473 . . .