Renin Gene Promoter Activity in Gc Cells Is Regulated by Camp and Thyroid-Hormone through Pit-1-Dependent Mechanisms

Gilbert, M. T., Sun, J. D., Yan, Y., Oddoux, C., Lazarus, A., Tansey, W. P., Lavin, T. N., Catanzaro, D. F. (November 1994) Renin Gene Promoter Activity in Gc Cells Is Regulated by Camp and Thyroid-Hormone through Pit-1-Dependent Mechanisms. Journal of Biological Chemistry, 269 (45). pp. 28049-28054. ISSN 0021-9258

URL: http://www.ncbi.nlm.nih.gov/pubmed/7961740

Abstract

Transcriptional activity of human renin gene (hREN) 5'-flanking DNA sequences in pituitary cells is highly dependent on binding of the pituitary-specific transcription factor Pit-1. Pit-1 has been implicated in cAMP regulation of a number of pituitary genes and has also been shown to interact with thyroid hormone (T-3) receptors in mediating T-3 responsiveness of the rat growth hormone gene. In the present study we examine the effects of forskolin and T-3 on the expression of luciferase hybrid genes containing hREN 5'-flanking DNAs (hREN.luc) transiently transfected into the pituitary cell line GC. Basal activities of all hREN.luc constructs transfected into cells grown in media containing serum stripped of hormones were low. Addition of forskolin stimulated expression up to 48 fold, depending on the hREN sequences present. The hREN sequence -148 to +18 was sufficient for both maximal expression and maximal stimulation by forskolin. Mutagenesis of the Pit-1 site between -82 and -58 reduced forskolin induction 4-5-fold. In addition to the Pit-1 site, the sequence between -148 and -98 was also required for maximal activity and forskolin induction. T-3 on its own had no effect on hREN promoter activity in GC cells, but suppressed the effects of forskolin. Gel mobility shift and Western blot analyses indicated that forskolin treatment had no effect on Pit-1 DNA binding or Pit-1 levels. However, T-3 reduced Pit-1 levels which was reflected in lower DNA binding under the conditions employed. Taken together, these findings emphasize the importance of cAMP-dependent mechanisms in directing renin gene expression.

Item Type: Paper
Uncontrolled Keywords: TRANSCRIPTION FACTOR PIT-1 RAT GROWTH-HORMONE THYROTROPIN-BETA-GENE HUMAN PROLACTIN GENE POU-DOMAIN ANGIOTENSIN SYSTEM RESPONSE ELEMENTS DNA-SEQUENCES EXPRESSION BINDING
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA expression > promoter
CSHL Authors:
Communities: CSHL labs > Tansey lab
Depositing User: Matt Covey
Date: November 1994
Date Deposited: 25 Aug 2015 18:48
Last Modified: 25 Aug 2015 18:48
Related URLs:
URI: http://repository.cshl.edu/id/eprint/31428

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