title: Isolation of marmoset P450c17 cDNA and gene regulation in vitro.
abstract: Marmosets express a fetal zone during adrenal development but fail to produce significant amounts of C19 steroids in adulthood. It is not clear however if P450c17 regulation is different from that in humans/rhesus or the primary sequence is altered. To this end we isolated marmoset and rhesus adrenocortical cells, and treated the cells with known regulators of P450c17 expression for 48 h. P450c17 protein increased with Forskolin (F) treatment, but was marginally inhibited by AII (A) and TPA (T) alone. Combined A + F and T + F dramatically ablated the F response. Cortisol levels (EIA) increased upon F treatment and were inhibited by A and T. Combination of treatments partially inhibited the F-induced response. The protein-coding region of marmoset and rhesus P450c17 cDNAs were then isolated from adrenals using RT-PCR/TA cloning. Marmoset P450c17 shows one amino acid deletion but otherwise shares 90.6% and 91.4% homologies with the human and rhesus cDNA sequences, and 82.4% and 85% homologies with the human and rhesus predicted AA sequences, respectively. Since marmoset adrenocortical cells exhibit similar endocrine function to rhesus, impaired 17,20-lyase activity in the adult marmoset adrenal may in part be due to differences in the primary sequence.
authors.affiliation: Perinatal Research Laboratories, Department of Obstetrics and Gynaecology, University of Wisconsin, Madison, Wisconsin 53715, USA.
authors.forename: J Christina
authors.initials: JC
authors.lastname: Pattison
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Conley
authors.forename: Ian M
authors.initials: IM
authors.lastname: Bird
mesh_terms: D000302:Adrenal Cortex / Q000166:cytology / Q000378:metabolism; D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D002144:Callithrix / Q000235:genetics*; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D018076:DNA, Complementary / Q000302:isolation & purification*; D004357:Drug Synergism; D005786:Gene Expression Regulation; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism; D008253:Macaca mulatta / Q000235:genetics; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D018076:DNA, Complementary; D000804:Angiotensin II; D005576:Colforsin; D013254:Steroid 17-alpha-Hydroxylase; D013755:Tetradecanoylphorbol Acetate; D006854:Hydrocortisone