Renin gene promoter activity in GC cells is regulated by cAMP and thyroid hormone through Pit-1-dependent mechanisms

Matthew T. Gilbert, Jidong Sun, Yan Yan, Carole Oddoux, Amy Lazarus, William P. Tansey, Thomas N. Lavin, Daniel F. Catanzaro

Research output: Contribution to journalArticle

23 Citations (Scopus)

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 (T3) receptors in mediating T3 responsiveness of the rat growth hormone gene. In the present study we examine the effects of forskolin and T3 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. T3 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, T3 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.

Original languageEnglish (US)
Pages (from-to)28049-28054
Number of pages6
JournalJournal of Biological Chemistry
Volume269
Issue number45
StatePublished - Nov 11 1994

Fingerprint

Thyroid Hormones
Renin
Genes
Colforsin
Thyroid Hormone Receptors
Human Activities
DNA
Transcription Factor Pit-1
Cells
Mutagenesis
5' Flanking Region
DNA sequences
Luciferases
Gene expression
Growth Hormone
Rats
Western Blotting
Gels
Hormones
Gene Expression

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Gilbert, M. T., Sun, J., Yan, Y., Oddoux, C., Lazarus, A., Tansey, W. P., ... Catanzaro, D. F. (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), 28049-28054.

Renin gene promoter activity in GC cells is regulated by cAMP and thyroid hormone through Pit-1-dependent mechanisms. / Gilbert, Matthew T.; Sun, Jidong; Yan, Yan; Oddoux, Carole; Lazarus, Amy; Tansey, William P.; Lavin, Thomas N.; Catanzaro, Daniel F.

In: Journal of Biological Chemistry, Vol. 269, No. 45, 11.11.1994, p. 28049-28054.

Research output: Contribution to journalArticle

Gilbert, MT, Sun, J, Yan, Y, Oddoux, C, Lazarus, A, Tansey, WP, Lavin, TN & Catanzaro, DF 1994, 'Renin gene promoter activity in GC cells is regulated by cAMP and thyroid hormone through Pit-1-dependent mechanisms', Journal of Biological Chemistry, vol. 269, no. 45, pp. 28049-28054.
Gilbert MT, Sun J, Yan Y, Oddoux C, Lazarus A, Tansey WP et al. Renin gene promoter activity in GC cells is regulated by cAMP and thyroid hormone through Pit-1-dependent mechanisms. Journal of Biological Chemistry. 1994 Nov 11;269(45):28049-28054.
Gilbert, Matthew T. ; Sun, Jidong ; Yan, Yan ; Oddoux, Carole ; Lazarus, Amy ; Tansey, William P. ; Lavin, Thomas N. ; Catanzaro, Daniel F. / Renin gene promoter activity in GC cells is regulated by cAMP and thyroid hormone through Pit-1-dependent mechanisms. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 45. pp. 28049-28054.
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