Thyrotropin-releasing hormone action on the prolactin promoter is mediated by the POU protein pit-1

• Published in: Molecular endocrinology (Baltimore, Md.) Vol. 5; no. 4; pp. 535 - 541
• Main Authors: Yan, G Z, Pan, W T, Bancroft, C
• Format: Journal Article
• Language: English
• Published: United States 01.04.1991
• Subjects: Animals; Base Sequence; Chromosome Mapping; DNA-Binding Proteins - physiology; Gene Expression Regulation; In Vitro Techniques; Molecular Sequence Data; Oligonucleotide Probes; Pituitary Gland - metabolism; Prolactin - biosynthesis; Promoter Regions, Genetic - drug effects; Rats; Thyrotropin-Releasing Hormone - pharmacology; Transcription Factor Pit-1; Transcription Factors - physiology; Transcription, Genetic - drug effects
• ISSN: 0888-8809; 1944-9917
• DOI: 10.1210/mend-5-4-535

TRH is known to regulate transcription of the PRL gene in pituitary cells, but little is known about the mechanism involved. We have characterized TRH response elements (TRHREs) in the promoter region of the rat PRL gene and the gene-proximal protein that transmits the TRH signal to these elements. Exposure of GH3 rat pituitary cells to TRH yielded a large specific stimulation of transient expression of a PRL-chloramphenicol acetyltransferase (PRL-CAT) construct containing the PRL promoter region [(-204)PRL-CAT]. Analysis of 5' deletions of this construct implied that regions -174/-113 and -75/+38 each contain a TRHRE. GH3 cell nuclear extracts are known to footprint four sites, termed, respectively, 1P-4P, on the PRL promoter region. The TRHRE between positions -75/+38 was identified as element 1P, residing at -63/-39, since two copies of a 1P oligodeoxynucleotide transferred a TRH response to either (-39)PRL-CAT or mouse metallothionein-CAT construct (-39)mMT-CAT. Similarly, the more proximal TRHRE may be element 3P, residing at -167/-144, since two copies of this element also transferred a TRH response to (-39)PRL-CAT. Binding of pit-1 to site 1P is known to be capable of activating pituitary cell-specific PRL gene expression. To investigate whether pit-1 can also transduce a TRH signal to this site, oligodeoxynucleotides were prepared corresponding to mutations in either or both of two consensus sequences in site 1P.