Promoter regulatory elements and DNase I-hypersensitive sites involved in serglycin proteoglycan gene expression in human erythroleukemia, CHRF 288-11, and HL-60 cells.
We have compared regulation of the serglycin gene in human erythroleukemia (HEL) and CHRF 288-11 cells, which have megakaryocytic characteristics, with promyelocytic HL-60 cells. Deletion constructs were prepared from the region -1123/+42 to -20/+42, and putative regulatory sites were mutated. In all three cell lines, the two major regulatory elements for constitutive expression were the (-80)ets site and the cyclic AMP response element (CRE) half-site at -70. A protein from HEL and CHRF, but not HL60, nuclear extracts bound to the (-80)ets site. Another protein from all three cell lines bound to the (-70)CRE. Phorbol 12-myristate 13-acetate (PMA) and dibutyryl cyclic AMP (dbcAMP) increased expression of the reporter in HEL cells 2.5-3- and 4.5-fold, respectively, from all constructs except those with (-70)CRE mutations. PMA virtually eliminated expression of serglycin mRNA and promoter constructs, but dbcAMP increased expression in HL-60 cells. The effects of PMA and dbcAMP on promoter expression correlated with mRNA expression. The strengths of two DNase I-hypersensitive sites in the 5'-flanking region and the first intron in all three cells correlated with relative endogenous serglycin mRNA expression. An additional DNase I-hypersensitive site in HL60 DNA in the first intron may be related to the high serglycin expression in HL60 relative to HEL or CHRF cells.
Schick
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BP
Cardeza Foundation for Hematologic Research, Jefferson Medical College of Thomas Jefferson University, 1015 Walnut Street, Philadelphia, PA 19107, USA. barbara.schick@mail.tju.edu
Petrushina
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Brodbeck
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Castronuevo
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D003994:Bucladesine / Q000494:pharmacology; D003850:Deoxyribonuclease I / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D018922:HL-60 Cells; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D007438:Introns; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D008533:Megakaryocytes / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011509:Proteoglycans / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D033921:Vesicular Transport Proteins
D011509:Proteoglycans; D012333:RNA, Messenger; D033921:Vesicular Transport Proteins; C075430:serglycin; D003994:Bucladesine; D003850:Deoxyribonuclease I; D013755:Tetradecanoylphorbol Acetate