Characterization of the promoter of the gene for a mouse vas deferens protein related to the aldo-keto reductase superfamily: effect of steroid hormones and phorbol esters.
Mouse vas deferens protein (MVDP) is a member of the aldo-keto reductase family regulated by androgens. The expression of a hybrid gene containing the promoter of the MVDP gene and the chloramphenicol acetyl transferase (CAT) gene coding region was analyzed in two cell lines that do not normally express the MVDP gene: T47D and CV1 cells. A small region of the promoter (-121 to +41) was able to direct significant expression of the reporter gene in both cell lines. Additional elements, between -510 and -121 modulate basal expression in a cell-dependent manner. Interestingly, the 162 bp fragment serves as an androgen-dependent enhancer, and mutation of the consensus ARE sequence located between positions -111 and -97 resulted in a loss of androgen response in both cell lines. Additional elements, upstream of the enhancer, modulate induction positively or negatively in relation to the cell line used. The expression of different MVDP-CAT constructs was more effectively induced by androgens than by glucocorticoids at physiological hormonal concentrations. In addition to the 162 bp enhancer, sequences upstream of -510 were also required for specific androgen regulation. Phorbol 12-myristate 13-acetate (TPA) had no effect on basal activity of the 1.8 kb MVDP-CAT construct but strongly enhanced the induction by androgens.
CNRS URA 1940, Université Blaise Pascal, Aubière, France.
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Fabre
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Darne
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Veyssiere
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Jean
D000429:Alcohol Oxidoreductases / Q000145:classification*; D000449:Aldehyde Reductase; D000728:Androgens / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002277:Carcinoma / Q000235:genetics / Q000473:pathology; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D013196:Dihydrotestosterone / Q000378:metabolism / Q000494:pharmacology; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D005938:Glucocorticoids / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology; D015674:Mammary Neoplasms, Animal / Q000235:genetics / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D011374:Progesterone / Q000378:metabolism / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D011817:Rabbits; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D000728:Androgens; D005938:Glucocorticoids; D011506:Proteins; D013196:Dihydrotestosterone; D011374:Progesterone; D000429:Alcohol Oxidoreductases; C066498:Akr1b7 protein, mouse; D000449:Aldehyde Reductase; D015500:Chloramphenicol O-Acetyltransferase; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate