0 16 Characterization characterization NN 17 19 of of IN 20 23 the the DT 24 30 murine murine JJ 31 47 cyclin-dependent cyclin-dependent JJ 48 54 kinase kinase NNP 55 64 inhibitor inhibitor NN 65 69 gene gene NN 70 77 p27Kip1 p27kip1 NN 77 78 . . . 80 83 The the DT 84 100 cyclin-dependent cyclin-dependent JJ 101 107 kinase kinase NNP 108 117 inhibitor inhibitor NN 118 125 p27Kip1 p27kip1 NN 126 131 plays play VBZ 132 134 an an DT 135 144 important important JJ 145 149 role role NN 150 152 in in IN 153 163 regulating regulate VBG 164 174 cell-cycle cell-cycle JJ 175 187 progression. progression. NN 188 195 p27Kip1 p27kip1 NN 196 204 directly directly RB 205 213 inhibits inhibit VBZ 214 217 the the DT 218 227 catalytic catalytic JJ 228 236 activity activity NN 237 239 of of IN 240 251 cyclin/cdks cyclin/cdk NNS 252 253 ( ( ( 253 269 cyclin-dependent cyclin-dependent JJ 270 276 kinase kinase NNP 276 277 ) ) ) 278 287 complexes complex NNS 288 294 and/or and/or CC 295 305 interferes interfere VBZ 306 316 physically physically RB 317 321 with with IN 322 333 cyclin/cdks cyclin/cdk NNS 334 344 activation activation NN 345 347 by by IN 348 351 CAK CAK NNP 351 352 . . . 353 366 Interestingly interestingly RB 366 367 , , , 368 371 the the DT 372 382 expression expression NN 383 388 level level NN 389 391 of of IN 392 399 p27Kip1 p27kip1 NN 400 404 mRNA mRNA NNP 405 408 was be VBD 409 416 maximal maximal JJ 417 419 in in IN 420 427 resting rest VBG 428 430 Go Go NNP 431 438 T-cells T-cell NNP 439 442 and and CC 443 450 rapidly rapidly RB 451 459 declined decline VBD 460 469 following follow VBG 470 478 anti-CD3 anti-cd3 JJ 479 489 activation activation NN 489 490 . . . 491 493 We we PRP 494 500 report report VBP 501 505 here here RB 506 509 the the DT 510 517 cloning cloning NN 518 520 of of IN 521 528 p27Kip1 p27kip1 NN 529 533 gene gene NN 534 538 from from IN 539 545 murine murine JJ 546 553 genomic genomic JJ 554 557 DNA dna NN 558 561 and and CC 562 565 the the DT 566 576 functional functional JJ 577 585 analysis analysis NN 586 588 of of IN 589 592 the the DT 593 601 promoter promoter NN 602 604 of of IN 605 608 the the DT 609 616 p27Kip1 p27kip1 NN 617 621 gene gene NN 621 622 . . . 623 626 The the DT 627 631 gene gene NN 632 640 consists consist VBZ 641 643 of of IN 644 646 at at IN 647 652 least least JJS 653 658 three three CD 659 664 exons exon NNS 665 668 and and CC 669 674 spans span NNS 675 679 more more JJR 680 684 than than IN 685 688 5.6 5.6 CD 689 691 kb kb NN 692 694 of of IN 695 698 DNA DNA NNP 698 699 . . . 700 706 Primer Primer NNP 707 716 extension extension NN 717 720 and and CC 721 729 nuclease nuclease NN 730 732 S1 s1 NN 733 743 protection protection NN 744 752 analysis analysis NN 753 761 revealed reveal VBD 762 765 two two CD 766 771 major major JJ 772 785 transcription transcription NN 786 796 initiation initiation NN 797 802 sites site NNS 802 803 . . . 804 807 The the DT 808 816 promoter promoter NN 817 823 region region NN 824 830 lacked lack VBD 831 832 a a DT 833 837 TATA TATA NNP 838 841 box box NN 842 845 but but CC 846 855 contained contain VBD 856 865 potential potential JJ 866 873 binding bind VBG 874 879 sites site NNS 880 883 for for IN 884 887 the the DT 888 903 transcriptional transcriptional JJ 904 911 factors factor NNS 912 921 including include VBG 922 925 two two CD 926 929 Sp1 sp1 NN 929 930 , , , 931 934 CRE CRE NNP 934 935 , , , 936 939 Myb Myb NNP 940 943 and and CC 944 948 NFkB NFkB NNP 949 956 located located JJ 957 959 at at IN 960 969 positions position NNS 970 974 -153 -153 CD 974 975 , , , 976 980 -178 -178 CD 980 981 , , , 982 986 -286 -286 CD 986 987 , , , 988 992 -875 -875 CD 992 993 , , , 994 997 and and CC 998 1003 -1011 -1011 CD 1003 1004 , , , 1005 1017 respectively respectively RB 1017 1018 . . . 1019 1021 To to TO 1022 1029 analyze analyze VB 1030 1033 the the DT 1034 1044 regulatory regulatory JJ 1045 1055 mechanisms mechanism NNS 1056 1067 controlling control VBG 1068 1075 p27Kip1 p27kip1 NN 1076 1080 gene gene NN 1081 1091 expression expression NN 1091 1092 , , , 1093 1095 we we PRP 1096 1109 characterized characterize VBD 1110 1113 the the DT 1114 1125 5’-flanking 5'-flanking JJ 1126 1132 region region NN 1133 1137 from from IN 1138 1140 nt nt NN 1141 1146 -1609 -1609 CD 1147 1149 to to TO 1150 1154 +178 +178 CD 1154 1155 . . . 1156 1159 The the DT 1160 1164 -326 -326 CD 1165 1167 to to TO 1168 1172 -615 -615 CD 1173 1179 region region NN 1180 1189 contained contain VBD 1190 1198 positive positive JJ 1199 1209 regulatory regulatory JJ 1210 1218 elements element NNS 1218 1219 . . .