Stimulation of the novel estrogen receptor-alpha intronic TERP-1 promoter by estrogens, androgen, pituitary adenylate cyclase-activating peptide, and forskolin, and autoregulation by TERP-1 protein.
The estrogen receptor-alpha (ERalpha) pituitary-specific variant, TERP-1, is regulated dramatically by physiological status. We examined hormonal regulation of the TERP-1 promoter in transient transfection assays in GH3 somatolactotrope cells. We found that 17beta-estradiol (E2), genistein, androgen, pituitary adenylate cyclase-activating peptide, and forskolin (FSK) all stimulated TERP-1 promoter activity, whereas progesterone had no effect. ERalpha bound to a palindromic estrogen response element (ERE) and two half-site EREs; mutation of any of these sites decreased basal expression and completely obliterated E2 stimulation. In contrast, mutation of an activator protein-1 site decreased basal and FSK-stimulated promoter activity, but not E2 or androgen stimulation. The pure antiestrogen ICI 182,780 suppressed E2 and genistein, but not FSK or androgen, stimulation. Similarly, mutation of the ERE palindrome or half-site EREs suppressed promoter stimulation by E2 and genistein, but not by androgen or FSK. Because TERP-1 levels regulate ERalpha function on model promoters, we tested TERP-1 modulation of its own and other physiological promoters. TERP-1 suppressed basal and E2-stimulated expression of its own promoter. TERP-1 suppression required the ERE regions of the promoter, and the dimerization domain of TERP-1. TERP-1 overexpression also suppressed E2 stimulation of the progesterone receptor and prolactin promoters. Thus, estrogens, androgen, and FSK can stimulate TERP-1 promoter activity, and increased TERP-1 expression modulates E2 stimulation of physiological promoters. These data suggest that TERP-1 regulation may play a significant role in modifying pituitary ERalpha responses.
Bryant
Winnifred M
WM
Division of Endocrinology and Metabolism, Department of Internal Medicine, University of Virginia, Charlottesville, Virginia 22903, USA.
Gibson
Matthew A
MA
Shupnik
Margaret A
MA
D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology*; D017931:DNA Primers; D015342:DNA Probes; D047628:Estrogen Receptor alpha / Q000235:genetics*; D004967:Estrogens / Q000494:pharmacology*; D007438:Introns / Q000187:drug effects / Q000235:genetics*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000502:physiology; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D051381:Rats; D011960:Receptors, Estrogen / Q000187:drug effects / Q000235:genetics*; D014162:Transfection
D000728:Androgens; D017931:DNA Primers; D015342:DNA Probes; D047628:Estrogen Receptor alpha; D004967:Estrogens; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D011960:Receptors, Estrogen; C118565:TERP-1; D005576:Colforsin