0 3 Sp1 sp1 NN 4 13 functions function NNS 14 16 in in IN 17 18 a a DT 19 38 chromatin-dependent chromatin-dependent JJ 39 45 manner manner NN 46 48 to to TO 49 56 augment augment VB 57 62 human human JJ 63 75 alpha-globin alpha-globin NN 76 84 promoter promoter NN 85 93 activity activity NN 93 94 . . . 96 99 The the DT 100 101 5 5 CD 101 102 ’ ' SYM 103 111 flanking flanking NN 112 118 region region NN 119 121 of of IN 122 125 the the DT 126 131 human human JJ 132 144 alpha-globin alpha-globin NN 145 149 gene gene NN 150 152 is be VBZ 153 159 highly highly RB 160 161 G g G 162 163 + + + 164 165 C C NNP 166 170 rich rich JJ 171 174 and and CC 175 183 contains contain VBZ 184 192 multiple multiple JJ 193 199 copies copy NNS 200 202 of of IN 203 206 the the DT 207 216 consensus consensus NN 217 225 sequence sequence NN 226 229 for for IN 230 233 the the DT 234 237 Sp1 sp1 NN 238 245 binding binding NN 246 250 site site NN 250 251 . . . 252 254 We we PRP 255 267 investigated investigate VBD 268 271 the the DT 272 276 role role NN 277 279 of of IN 280 284 this this DT 285 286 G g G 287 288 + + + 289 295 C-rich c-rich JJ 296 302 region region NN 303 305 in in IN 306 316 augmenting augment VBG 317 329 alpha-globin alpha-globin NN 330 338 promoter promoter NN 339 347 activity activity NN 348 350 in in IN 351 354 the the DT 355 363 presence presence NN 364 366 of of IN 367 370 the the DT 371 383 far-upstream far-upstream JJ 384 396 alpha-globin alpha-globin NN 397 405 enhancer enhancer NN 405 406 , , , 407 412 HS-40 HS-40 NNP 412 413 . . . 414 416 We we PRP 417 421 show show VBP 422 426 that that IN 427 429 in in IN 430 441 transiently transiently RB 442 453 transfected transfecte VBN 454 463 erythroid erythroid JJ 464 469 cells cell NNS 469 470 , , , 471 479 deletion deletion NN 480 482 of of IN 483 486 the the DT 487 499 alpha-globin alpha-globin NN 500 501 G g G 502 503 + + + 504 510 C-rich c-rich JJ 511 512 5 5 CD 512 513 ’ ' SYM 514 522 flanking flanking NN 523 529 region region NN 530 533 has have VBZ 534 536 no no DT 537 543 effect effect NN 544 546 on on IN 547 559 alpha-globin alpha-globin NN 560 568 promoter promoter NN 569 577 activity activity NN 577 578 . . . 579 586 However however RB 586 587 , , , 588 592 upon upon IN 593 599 stable stable JJ 600 611 integration integration NN 612 616 into into IN 617 626 chromatin chromatin NN 626 627 , , , 628 636 deletion deletion NN 637 639 of of IN 640 644 this this DT 645 651 region region NN 652 658 causes cause VBZ 659 660 a a DT 661 667 nearly nearly RB 668 670 90 90 CD 670 671 % % NN 672 680 decrease decrease NN 681 683 in in IN 684 692 promoter promoter NN 693 701 activity activity NN 702 710 compared compare VBN 711 715 with with IN 716 726 expression expression NN 727 731 from from IN 732 734 an an DT 735 747 alpha-globin alpha-globin NN 748 756 promoter promoter NN 757 766 retaining retain VBG 767 771 this this DT 772 778 region region NN 778 779 . . . 780 785 These these DT 786 793 results result NNS 794 801 suggest suggest VBP 802 806 that that IN 807 810 the the DT 811 823 alpha-globin alpha-globin NN 824 825 G g G 826 827 + + + 828 834 C-rich c-rich JJ 835 836 5 5 CD 836 837 ’ ' SYM 838 846 flanking flanking NN 847 853 region region NN 854 862 augments augment VBZ 863 875 alpha-globin alpha-globin NN 876 884 promoter promoter NN 885 893 activity activity NN 894 896 in in IN 897 898 a a DT 899 918 chromatin-dependent chromatin-dependent JJ 919 925 manner manner NN 925 926 . . . 927 929 We we PRP 930 937 further further RB 938 942 show show VBP 943 947 that that IN 948 952 this this DT 953 954 G g G 955 956 + + + 957 963 C-rich c-rich JJ 964 970 region region NN 971 973 is be VBZ 974 982 required require VBN 983 986 for for IN 987 990 the the DT 991 1001 activation activation NN 1002 1004 of of IN 1005 1017 alpha-globin alpha-globin NN 1018 1022 gene gene NN 1023 1033 expression expression NN 1034 1040 during during IN 1041 1050 erythroid erythroid JJ 1051 1066 differentiation differentiation NN 1066 1067 . . . 1068 1075 Finally finally RB 1075 1076 , , , 1077 1079 we we PRP 1080 1084 show show VBP 1085 1087 by by IN 1088 1092 both both CC 1093 1102 footprint footprint NN 1103 1111 analysis analysis NN 1112 1115 and and CC 1116 1126 functional functional JJ 1127 1133 assays assay NNS 1134 1138 that that IN 1139 1142 the the DT 1143 1150 ability ability NN 1151 1153 of of IN 1154 1157 the the DT 1158 1159 G g G 1160 1161 + + + 1162 1168 C-rich c-rich JJ 1169 1175 region region NN 1176 1178 to to TO 1179 1187 increase increase VB 1188 1200 alpha-globin alpha-globin NN 1201 1209 promoter promoter NN 1210 1218 activity activity NN 1219 1223 from from IN 1224 1225 a a DT 1226 1232 stably stably RB 1233 1243 integrated integrate VBN 1244 1256 alpha-globin alpha-globin NN 1257 1261 gene gene NN 1262 1264 is be VBZ 1265 1273 mediated mediate VBN 1274 1276 by by IN 1277 1280 its its PRP$ 1281 1289 multiple multiple JJ 1290 1297 binding bind VBG 1298 1303 sites site NNS 1304 1307 for for IN 1308 1311 the the DT 1312 1325 transcription transcription NN 1326 1332 factor factor NN 1333 1336 Sp1 sp1 NN 1336 1337 . . .