Protein phosphatase 2A and phosphoprotein SET regulate androgen production by P450c17.
Cytochrome P450c17 catalyzes 17 alpha-hydroxylation needed for cortisol synthesis and 17,20 lyase activity needed to produce sex steroids. Serine phosphorylation of P450c17 specifically increases 17,20 lyase activity, but the physiological factors regulating this effect remain unknown. Treating human adrenal NCI-H295A cells with the phosphatase inhibitors okadaic acid, fostriecin, and cantharidin increased 17,20 lyase activity, suggesting involvement of protein phosphatase 2A (PP2A) or 4 (PP4). PP2A but not PP4 inhibited 17,20 lyase activity in microsomes from cultured cells, but neither affected 17 alpha-hydroxylation. Inhibition of 17,20 lyase activity by PP2A was concentration-dependent, could be inhibited by okadaic acid, and was restored by endogenous protein kinases. PP2A but not PP4 coimmunoprecipitated with P450c17, and suppression of PP2A by small interfering RNA increased 17,20 lyase activity. Phosphoprotein SET found in adrenals inhibited PP2A, but not PP4, and fostered 17,20 lyase activity. The identification of PP2A and SET as post-translational regulators of androgen biosynthesis suggests potential additional mechanisms contributing to adrenarche and hyperandrogenic disorders such as polycystic ovary syndrome.
Pandey
Amit V
AV
Department of Pediatrics, University of California, San Francisco, California 94143-0978, USA.
Mellon
Synthia H
SH
Miller
Walter L
WL
D000306:Adrenal Cortex Neoplasms; D000728:Androgens / Q000096:biosynthesis*; D001483:Base Sequence; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D056488:Histone Chaperones; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology; D019319:Okadaic Acid / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D054648:Protein Phosphatase 2; D011506:Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors; D014407:Tumor Cells, Cultured
D000728:Androgens; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D056488:Histone Chaperones; D011506:Proteins; D011994:Recombinant Proteins; C513473:SET protein, human; D014157:Transcription Factors; D010768:Phosphoserine; D019319:Okadaic Acid; D013254:Steroid 17-alpha-Hydroxylase; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2