0 7 Cloning cloning NN 8 10 of of IN 11 14 the the DT 15 20 novel novel JJ 21 26 human human JJ 27 48 myeloid-cell-specific myeloid-cell-specific JJ 49 62 C/EBP-epsilon C/EBP-epsilon NNP 63 76 transcription transcription NN 77 83 factor factor NN 83 84 . . . 86 93 Chicken Chicken NNP 94 98 NF-M NF-M NNP 99 112 transcription transcription NN 113 119 factor factor NN 119 120 , , , 121 123 in in IN 124 135 cooperation cooperation NN 136 140 with with IN 141 147 either either CC 148 153 c-Myb c-myb NN 154 156 or or CC 157 162 v-Myb v-myb NN 162 163 , , , 164 166 is be VBZ 167 173 active active JJ 174 176 in in IN 177 180 the the DT 181 194 combinatorial combinatorial JJ 195 205 activation activation NN 206 208 of of IN 209 230 myeloid-cell-specific myeloid-cell-specific JJ 231 236 genes gene NNS 237 239 in in IN 240 252 heterologous heterologous JJ 253 257 cell cell NN 258 263 types type NNS 263 264 , , , 265 269 such such JJ 270 272 as as IN 273 282 embryonic embryonic JJ 283 294 fibroblasts fibroblast NNS 294 295 . . . 296 298 In in IN 299 305 humans human NNS 305 306 , , , 307 314 similar similar JJ 315 322 effects effect NNS 323 327 were be VBD 328 336 observed observe VBN 337 341 with with IN 342 352 homologous homologous JJ 353 360 members member NNS 361 363 of of IN 364 367 the the DT 368 390 CCAAT/enhancer-binding ccaat/enhancer-binding JJ 391 398 protein protein NN 399 400 ( ( ( 400 405 C/EBP C/EBP NNP 405 406 ) ) ) 407 413 family family NN 414 416 of of IN 417 432 transcriptional transcriptional JJ 433 443 regulators regulator NNS 443 444 , , , 445 455 especially especially RB 456 459 the the DT 460 465 human human JJ 466 473 homolog homolog NN 474 476 of of IN 477 484 chicken chicken NN 485 489 NF-M NF-M NNP 489 490 , , , 491 501 C/EBP-beta C/EBP-beta NNP 502 503 ( ( ( 503 509 NF-IL6 NF-IL6 NNP 509 510 ) ) ) 510 511 . . . 512 519 However however RB 519 520 , , , 521 524 the the DT 525 531 NF-IL6 NF-IL6 NNP 532 536 gene gene NN 537 539 is be VBZ 540 549 expressed express VBN 550 552 in in IN 553 554 a a DT 555 562 variety variety NN 563 565 of of IN 566 576 nonmyeloid nonmyeloid JJ 577 581 cell cell NN 582 587 types type NNS 588 591 and and CC 592 594 is be VBZ 595 603 strongly strongly RB 604 613 inducible inducible JJ 614 616 in in IN 617 625 response response NN 626 628 to to TO 629 641 inflammatory inflammatory JJ 642 649 stimuli stimulus NNS 649 650 , , , 651 657 making make VBG 658 660 it it PRP 661 663 an an DT 664 672 unlikely unlikely JJ 673 682 candidate candidate NN 683 685 to to TO 686 690 have have VB 691 693 an an DT 694 703 exclusive exclusive JJ 704 708 role role NN 709 711 as as IN 712 713 a a DT 714 727 combinatorial combinatorial JJ 728 743 differentiation differentiation NN 744 750 switch switch NN 751 757 during during IN 758 770 myelopoiesis myelopoiesis NN 771 773 in in IN 774 779 human human JJ 780 785 cells cell NNS 785 786 . . . 787 789 By by IN 790 795 using use VBG 796 797 a a DT 798 805 reverse reverse JJ 806 829 transcription-PCR-based transcription-pcr-based JJ 830 838 approach approach NN 839 842 and and CC 843 844 a a DT 845 848 set set NN 849 851 of of IN 852 859 primers primer NNS 860 868 specific specific JJ 869 872 for for IN 873 876 the the DT 877 888 DNA-binding dna-binding JJ 889 896 domains domain NNS 897 899 of of IN 900 906 highly highly RB 907 917 homologous homologous JJ 918 925 members member NNS 926 928 of of IN 929 932 the the DT 933 938 C/EBP C/EBP NNP 939 945 family family NN 946 948 of of IN 949 964 transcriptional transcriptional JJ 965 975 regulators regulator NNS 975 976 , , , 977 979 we we PRP 980 984 have have VBP 985 991 cloned clone VBN 992 993 a a DT 994 999 novel novel JJ 1000 1005 human human JJ 1006 1010 gene gene NN 1011 1019 encoding encode VBG 1020 1021 a a DT 1022 1028 member member NN 1029 1031 of of IN 1032 1035 the the DT 1036 1041 C/EBP C/EBP NNP 1042 1046 gene gene NN 1047 1053 family family NN 1053 1054 , , , 1055 1065 identified identify VBN 1066 1068 as as IN 1069 1072 the the DT 1073 1078 human human JJ 1079 1086 homolog homolog NN 1087 1089 of of IN 1090 1094 CRP1 CRP1 NNP 1094 1095 , , , 1096 1109 C/EBP-epsilon C/EBP-epsilon NNP 1109 1110 . . . 1111 1112 A a DT 1113 1119 1.2-kb 1.2-kb JJ 1120 1124 cDNA cdna NN 1125 1133 encoding encode VBG 1134 1145 full-length full-length JJ 1146 1151 human human JJ 1152 1165 C/EBP-epsilon c/ebp-epsilon NN 1166 1169 was be VBD 1170 1176 cloned clone VBN 1177 1181 from from IN 1182 1183 a a DT 1184 1201 promyelocyte-late promyelocyte-late JJ 1202 1220 myeloblast-derived myeloblast-derived JJ 1221 1227 lambda lambda NN 1228 1232 gt11 gt11 NN 1233 1240 library library NN 1240 1241 . . . 1242 1251 Molecular molecular JJ 1252 1260 analysis analysis NN 1261 1263 of of IN 1264 1267 the the DT 1268 1272 cDNA cdna NN 1273 1276 and and CC 1277 1284 genomic genomic JJ 1285 1291 clones clone NNS 1292 1301 indicated indicate VBD 1302 1305 the the DT 1306 1314 presence presence NN 1315 1317 of of IN 1318 1321 two two CD 1322 1327 exons exon NNS 1328 1336 encoding encode VBG 1337 1338 a a DT 1339 1346 protein protein NN 1347 1351 with with IN 1352 1354 an an DT 1355 1363 apparent apparent JJ 1364 1373 molecular molecular JJ 1374 1378 mass mass NN 1379 1381 of of IN 1382 1384 32 32 CD 1385 1388 kDa kda NN 1389 1392 and and CC 1393 1394 a a DT 1395 1397 pI pi NN 1398 1400 of of IN 1401 1404 9.5 9.5 CD 1404 1405 . . . 1406 1412 Primer Primer NNP 1413 1422 extension extension NN 1423 1431 analysis analysis NN 1432 1434 of of IN 1435 1448 C/EBP-epsilon C/EBP-epsilon NNP 1449 1453 mRNA mRNA NNP 1454 1462 detected detect VBD 1463 1464 a a DT 1465 1471 single single JJ 1472 1477 major major JJ 1478 1491 transcription transcription NN 1492 1497 start start NN 1498 1502 site site NN 1503 1516 approximately approximately RB 1517 1520 200 200 CD 1521 1523 bp bp NN 1524 1532 upstream upstream RB 1533 1535 of of IN 1536 1539 the the DT 1540 1545 start start NN 1546 1551 codon codon NN 1551 1552 . . . 1553 1556 The the DT 1557 1565 putative putative JJ 1566 1574 promoter promoter NN 1575 1579 area area NN 1580 1582 is be VBZ 1583 1590 similar similar JJ 1591 1593 to to TO 1594 1599 those those DT 1600 1602 of of IN 1603 1610 several several JJ 1611 1616 other other JJ 1617 1638 myeloid-cell-specific myeloid-cell-specific JJ 1639 1644 genes gene NNS 1645 1647 in in IN 1648 1652 that that IN 1653 1655 it it PRP 1656 1664 contains contain VBZ 1665 1667 no no DT 1668 1674 TATAAA TATAAA NNP 1675 1678 box box NN 1679 1682 but but CC 1683 1686 has have VBZ 1687 1688 a a DT 1689 1695 number number NN 1696 1698 of of IN 1699 1710 purine-rich purine-rich JJ 1711 1720 stretches stretch NNS 1721 1725 with with IN 1726 1734 multiple multiple JJ 1735 1740 sites site NNS 1741 1744 for for IN 1745 1748 the the DT 1749 1756 factors factor NNS 1757 1759 of of IN 1760 1763 the the DT 1764 1767 Ets Ets NNP 1768 1774 family family NN 1775 1777 of of IN 1778 1793 transcriptional transcriptional JJ 1794 1804 regulators regulator NNS 1804 1805 . . . 1806 1814 Northern Northern NNP 1815 1819 blot blot NN 1820 1828 analyses analysis NNS 1829 1838 indicated indicate VBD 1839 1840 a a DT 1841 1847 highly highly RB 1848 1858 restricted restricted JJ 1859 1863 mRNA mrna NN 1864 1874 expression expression NN 1875 1882 pattern pattern NN 1882 1883 , , , 1884 1888 with with IN 1889 1892 the the DT 1893 1902 strongest strong JJS 1903 1913 expression expression NN 1914 1923 occurring occur VBG 1924 1926 in in IN 1927 1939 promyelocyte promyelocyte NN 1940 1943 and and CC 1944 1964 late-myeloblast-like late-myeloblast-like JJ 1965 1969 cell cell NN 1970 1975 lines line NNS 1975 1976 . . . 1977 1984 Western western NN 1985 1989 blot blot NN 1990 1993 and and CC 1994 2013 immunoprecipitation immunoprecipitation NN 2014 2021 studies study NNS 2022 2027 using use VBG 2028 2034 rabbit rabbit NN 2035 2053 anti-C/EBP-epsilon anti-c/ebp-epsilon JJ 2054 2064 antibodies antibody NNS 2065 2071 raised raise VBN 2072 2079 against against IN 2080 2083 the the DT 2084 2094 N-terminal n-terminal JJ 2095 2102 portion portion NN 2103 2105 of of IN 2106 2119 C/EBP-epsilon C/EBP-epsilon NNP 2120 2121 ( ( ( 2121 2126 amino amino NN 2127 2132 acids acid NNS 2133 2134 1 1 CD 2135 2137 to to TO 2138 2141 115 115 CD 2141 2142 ) ) ) 2143 2149 showed show VBD 2150 2154 that that IN 2155 2168 C/EBP-epsilon C/EBP-epsilon NNP 2169 2171 is be VBZ 2172 2173 a a DT 2174 2180 32-kDa 32-kda JJ 2181 2188 nuclear nuclear JJ 2189 2203 phosphoprotein phosphoprotein NN 2203 2204 . . . 2205 2208 The the DT 2209 2214 human human JJ 2215 2228 C/EBP-epsilon C/EBP-epsilon NNP 2229 2236 protein protein NN 2237 2246 exhibited exhibit VBD 2247 2253 strong strong JJ 2254 2257 and and CC 2258 2266 specific specific JJ 2267 2274 binding binding NN 2275 2277 to to TO 2278 2293 double-stranded double-stranded JJ 2294 2297 DNA dna NN 2298 2308 containing contain VBG 2309 2318 consensus consensus NN 2319 2324 C/EBP C/EBP NNP 2325 2330 sites site NNS 2330 2331 . . . 2332 2346 Cotransfection cotransfection NN 2347 2349 of of IN 2350 2353 the the DT 2354 2367 C/EBP-epsilon C/EBP-epsilon NNP 2368 2373 sense sense NN 2374 2377 and and CC 2378 2387 antisense antisense JJ 2388 2398 expression expression NN 2399 2409 constructs construct NNS 2410 2418 together together RB 2419 2423 with with IN 2424 2439 chloramphenicol chloramphenicol NN 2440 2457 acetyltransferase acetyltransferase NN 2458 2466 reporter reporter NN 2467 2474 vectors vector NNS 2475 2485 containing contain VBG 2486 2507 myeloid-cell-specific myeloid-cell-specific JJ 2508 2513 c-mim c-mim NN 2514 2517 and and CC 2518 2523 human human JJ 2524 2539 myeloperoxidase myeloperoxidase NN 2540 2549 promoters promoter NNS 2550 2559 suggested suggest VBD 2560 2561 a a DT 2562 2566 role role NN 2567 2570 for for IN 2571 2584 C/EBP-epsilon C/EBP-epsilon NNP 2585 2598 transcription transcription NN 2599 2605 factor factor NN 2606 2608 in in IN 2609 2612 the the DT 2613 2623 regulation regulation NN 2624 2626 of of IN 2627 2628 a a DT 2629 2635 subset subset NN 2636 2638 of of IN 2639 2660 myeloid-cell-specific myeloid-cell-specific JJ 2661 2666 genes gene NNS 2666 2667 . . . 2668 2677 Transient transient JJ 2678 2689 tranfection tranfection NN 2690 2692 of of IN 2693 2694 a a DT 2695 2707 promyelocyte promyelocyte JJ 2708 2712 cell cell NN 2713 2717 line line NN 2718 2719 ( ( ( 2719 2722 NB4 nb4 NN 2722 2723 ) ) ) 2724 2728 with with IN 2729 2730 a a DT 2731 2744 C/EBP-epsilon C/EBP-epsilon NNP 2745 2755 expression expression NN 2756 2763 plasmid plasmid NN 2764 2773 increased increase VBD 2774 2778 cell cell NN 2779 2785 growth growth NN 2786 2788 by by IN 2789 2798 sevenfold sevenfold RB 2798 2799 , , , 2800 2805 while while IN 2806 2815 antisense antisense JJ 2816 2829 C/EBP-epsilon C/EBP-epsilon NNP 2830 2836 caused cause VBD 2837 2838 a a DT 2839 2847 fivefold fivefold JJ 2848 2856 decrease decrease NN 2857 2859 in in IN 2860 2866 clonal clonal JJ 2867 2873 growth growth NN 2874 2876 of of IN 2877 2882 these these DT 2883 2888 cells cell NNS 2888 2889 . . .