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This version published online on February 24, 2005
Endocrinology, doi:10.1210/en.2004-1673
A more recent version of this article appeared on June 1, 2005
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Submitted on December 29, 2004
Accepted on February 16, 2005

Cloning and characterization of a 5' regulatory region of the PRAP/17{beta} HSD7 gene

Michael Risk, Aurora Shehu, Jifang Mao, Carlos O. Stocco, Laura T. Goldsmith, Jennifer M. Bowen-Shauver, and Geula Gibori*

Department of Physiology and Biophysics, University of Illinois at Chicago, 835 S. Wolcott (M/C 901), Chicago, IL 60612; Department of Obstetrics, Gynecology & Women's Health; New Jersey Medical School, 185 South Orange Avenue, PO Box 1709 Newark, NJ 07101-1709

* To whom correspondence should be addressed. E-mail: ggibori{at}uic.edu.

Prolactin receptor associated protein (PRAP) originally cloned in our laboratory was shown to be a novel, luteal isoform of 17{beta}hydroxysteroid dehydrogenase (17{beta}HSD7). In this study, we cloned the promoter region of rat PRAP/17{beta}HSD7 and investigated the mechanisms regulating both basal activity and LH-induced repression of this promoter. Truncated and site- specific mutants of PRAP/17{beta}HSD7 promoter identified two enhancer regions that contained highly conserved Sp1 binding site and bound Sp1 from nuclear extracts of both corpora lutea and a rat luteal cell line. Repression of PRAP/17{beta}HSD7 expression and promoter activity by human chorionic gonadotropin (hCG)/forskolin was localized to a -52 bp proximal segment of the promoter. This region contained a conserved CCAAT site and bound NF-Y; binding of this transcription factor was inhibited by hCG in vivo. Further, mutation of the NF-Y site in the -52 bp promoter-reporter construct abolished forskolin-mediated inhibition of the promoter in a rat luteal cell line. In summary, we have identified the promoter elements involved in the basal expression of PRAP/17{beta}HSD7. We have also found that LH-mediated repression of this gene is at the level of transcription and involves inhibition of NF-YA binding to the CCAAT site within the proximal promoter.
Key words: Ovary • Corpus luteum • steroidogenesis • hydroxysteroid dehydrogenase




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