Promoter regulatory elements and DNase I-hypersensitive sites involved in serglycin proteoglycan gene expression in human erythroleukemia, CHRF 288-11, and HL-60 cells.

Abstract

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.

• Authors: B P Schick, I Petrushina, K C Brodbeck, P Castronuevo
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 276 Issue 27 Pg. 24726-35 (Jul 06 2001) ISSN: 0021-9258 [Print] United States
• PMID: 11333275 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Proteoglycans
• RNA, Messenger
• Vesicular Transport Proteins
• serglycin
• Bucladesine
• Deoxyribonuclease I
• Tetradecanoylphorbol Acetate

Topics

• Bucladesine (pharmacology)
• Deoxyribonuclease I (metabolism)
• Gene Expression Regulation, Neoplastic
• HL-60 Cells
• Helix-Loop-Helix Motifs
• Humans
• Introns
• Leukemia, Erythroblastic, Acute (metabolism)
• Megakaryocytes (metabolism)
• Promoter Regions, Genetic
• Proteoglycans (genetics)
• RNA, Messenger (metabolism)
• Tetradecanoylphorbol Acetate (pharmacology)
• Vesicular Transport Proteins