0 15 Transcriptional transcriptional JJ 16 26 regulation regulation NN 27 29 of of IN 30 33 the the DT 34 42 vacuolar vacuolar JJ 43 54 H(+)-ATPase h(+)-atpase NN 55 57 B2 b2 NN 58 65 subunit subunit NN 66 70 gene gene NN 71 73 in in IN 74 89 differentiating differentiate VBG 90 95 THP-1 thp-1 NN 96 101 cells cell NNS 101 102 . . . 104 123 Monocyte-macrophage monocyte-macrophage JJ 124 139 differentiation differentiation NN 140 143 was be VBD 144 148 used use VBN 149 151 as as IN 152 153 a a DT 154 159 model model NN 160 166 system system NN 167 170 for for IN 171 179 studying study VBG 180 184 gene gene NN 185 195 regulation regulation NN 196 198 of of IN 199 202 the the DT 203 208 human human JJ 209 217 vacuolar vacuolar JJ 218 229 H(+)-ATPase h(+)-atpase NN 230 231 ( ( ( 231 239 V-ATPase v-atpase NN 239 240 ) ) ) 240 241 . . . 242 244 We we PRP 245 253 examined examine VBD 254 258 mRNA mrna NN 259 265 levels level NNS 266 268 of of IN 269 276 various various JJ 277 285 V-ATPase v-atpase NN 286 294 subunits subunit NNS 295 301 during during IN 302 317 differentiation differentiation NN 318 320 of of IN 321 325 both both DT 326 332 native native JJ 333 342 monocytes monocyte NNS 343 346 and and CC 347 350 the the DT 351 355 cell cell NN 356 360 line line NN 361 366 THP-1 thp-1 NN 366 367 , , , 368 371 and and CC 372 377 found find VBD 378 382 that that IN 383 398 transcriptional transcriptional JJ 399 402 and and CC 403 423 post-transcriptional post-transcriptional JJ 424 434 mechanisms mechanism NNS 435 440 could could MD 441 448 account account VB 449 452 for for IN 453 462 increases increase NNS 463 465 in in IN 466 470 cell cell NN 471 479 V-ATPase v-atpase NN 480 487 content content NN 487 488 . . . 489 493 From from IN 494 501 nuclear nuclear JJ 502 508 runoff runoff NN 509 520 experiments experiment NNS 520 521 , , , 522 524 we we PRP 525 530 found find VBD 531 535 that that IN 536 539 one one CD 540 547 subunit subunit NN 548 550 in in IN 551 561 particular particular JJ 561 562 , , , 563 566 the the DT 567 569 B2 b2 NN 570 577 isoform isoform NN 578 579 ( ( ( 579 581 Mr Mr NNP 582 583 = = JJ 584 590 56,000 56,000 CD 590 591 ) ) ) 591 592 , , , 593 596 was be VBD 597 606 amplified amplify VBN 607 616 primarily primarily RB 617 619 by by IN 620 635 transcriptional transcriptional JJ 636 641 means mean NNS 641 642 . . . 643 645 We we PRP 646 650 have have VBP 651 656 begun begin VBN 657 659 to to TO 660 667 examine examine VB 668 671 the the DT 672 681 structure structure NN 682 684 of of IN 685 688 the the DT 689 691 B2 b2 NN 692 699 subunit subunit NN 700 708 promoter promoter NN 709 715 region region NN 715 716 . . . 717 726 Isolation isolation NN 727 730 and and CC 731 741 sequencing sequencing NN 742 744 of of IN 745 748 the the DT 749 754 first first JJ 755 759 exon exon NN 760 763 and and CC 764 775 5’-flanking 5'-flanking JJ 776 782 region region NN 783 785 of of IN 786 790 this this DT 791 795 gene gene NN 796 802 reveal reveal VBP 803 804 a a DT 805 814 TATA-less tata-less JJ 815 823 promoter promoter NN 824 828 with with IN 829 830 a a DT 831 835 high high JJ 836 837 G G NNP 838 839 + + SYM 840 841 C C NNP 842 849 content content NN 849 850 . . . 851 857 Primer Primer NNP 858 867 extension extension NN 868 871 and and CC 872 884 ribonuclease ribonuclease NN 885 895 protection protection NN 896 904 analyses analysis NNS 905 913 indicate indicate VBP 914 915 a a DT 916 922 single single JJ 923 928 major major JJ 929 944 transcriptional transcriptional JJ 945 950 start start NN 951 955 site site NN 955 956 . . . 957 959 We we PRP 960 971 transfected transfecte VBD 972 991 promoter-luciferase promoter-luciferase JJ 992 1000 reporter reporter NN 1001 1009 plasmids plasmid NNS 1010 1014 into into IN 1015 1020 THP-1 thp-1 NN 1021 1026 cells cell NNS 1027 1029 to to TO 1030 1036 define define VB 1037 1046 sequences sequence NNS 1047 1051 that that WDT 1052 1059 mediate mediate VBP 1060 1075 transcriptional transcriptional JJ 1076 1083 control control NN 1084 1090 during during IN 1091 1099 monocyte monocyte NN 1100 1115 differentiation differentiation NN 1115 1116 . . . 1117 1119 We we PRP 1120 1125 found find VBD 1126 1130 that that IN 1131 1140 sequences sequence NNS 1141 1151 downstream downstream RB 1152 1156 from from IN 1157 1160 the the DT 1161 1176 transcriptional transcriptional JJ 1177 1182 start start NN 1183 1187 site site NN 1188 1192 were be VBD 1193 1203 sufficient sufficient JJ 1204 1206 to to TO 1207 1213 confer confer VB 1214 1223 increased increase VBN 1224 1234 expression expression NN 1235 1241 during during IN 1242 1247 THP-1 thp-1 NN 1248 1263 differentiation differentiation NN 1263 1264 . . . 1265 1270 DNase DNase NNP 1271 1272 I i CD 1273 1285 footprinting footprinting NN 1286 1289 and and CC 1290 1298 sequence sequence NN 1299 1307 analysis analysis NN 1308 1316 revealed reveal VBD 1317 1320 the the DT 1321 1330 existence existence NN 1331 1333 of of IN 1334 1342 multiple multiple JJ 1343 1346 AP2 ap2 NN 1347 1350 and and CC 1351 1354 Sp1 sp1 NN 1355 1362 binding binding NN 1363 1368 sites site NNS 1369 1371 in in IN 1372 1375 the the DT 1376 1391 5’-untranslated 5'-untranslated JJ 1392 1395 and and CC 1396 1404 proximal proximal JJ 1405 1411 coding coding NN 1412 1419 regions region NNS 1419 1420 . . .