Endocrinology, Vol 136, 5430-5437, Copyright © 1995 by Endocrine Society

ARTICLES

Cis-regulatory elements conferring cyclic 3',5'-adenosine monophosphate responsiveness of the progesterone receptor gene in transfected rat granulosa cells

OK Park-Sarge and KD Sarge
Department of Physiology, University of Kentucky, Lexington 40536-0084, USA.

We have previously shown that both pituitary gonadotropins and forskolin induce progesterone receptor (PR) messenger RNA expression at the level of transcription in granulosa cells of the rat ovary. To determine the DNA regulatory elements that are important for cAMP- induced transcription of the PR gene in the ovary, we examined the cAMP- induced activity of promoter sequences in rat granulosa cells transfected with various fusion constructs containing PRB promoter sequences linked to the luciferase reporter gene. When cells were transfected with a luciferase fusion construct containing the 1375-base pair 5'-flanking region of the rat PRB gene, forskolin treatment substantially increased luciferase activity. Analysis of a series of 5'- deletion mutants indicated that a minimal PRB promoter containing 116 base pairs of upstream sequence (-116/3) was sufficient to increase luciferase activity in response to forskolin in transfected rat granulosa cells. This promoter contains a consensus CCAAT site in reverse orientation (5'-ATTGG-3') and a consensus GC box (5'-GGGGCGGGCC- 3'), but no known cAMP-responsive element. Site-specific mutation of the GC box notably decreased both basal and cAMP-induced activity of this minimal PRB promoter. In addition, site-specific mutation of the CCAAT binding site within this proximal promoter of the PRB gene substantially decreased cAMP-induced activity, but did not significantly affect the basal activity of this promoter. Either mutation alone failed to abolish cAMP inducibility. In contrast, double mutation of both the GC box and the CCAAT box completely abolished cAMP inducibility, suggesting that the GC box and the CCAAT box act together to mediate cAMP-induced transcription of the PRB gene. Gel shift analysis shows that the minimal PRB promoter sequences form multiple complexes with nuclear proteins of granulosa cells, all of which are specifically competed by oligonucleotides containing the GC box and the CCAAT box. Taken together, our results suggest a functional role for transcription factors binding the GC box and the CCAAT box in mediating cAMP-induced transcription of the rat PRB promoter in rat granulosa cells.

This article has been cited by other articles:

				M. Lindeberg, K. Carlstrom, O. Ritvos, and O. Hovatta Gonadotrophin stimulation of non-luteinized granulosa cells increases steroid production and the expression of enzymes involved in estrogen and progesterone synthesis Hum. Reprod., February 1, 2007; 22(2): 401 - 406. [Abstract] [Full Text] [PDF]

				M. Hsieh, S. M. Mulders, R. R. Friis, A. Dharmarajan, and J. S. Richards Expression and Localization of Secreted Frizzled-Related Protein-4 in the Rodent Ovary: Evidence for Selective Up-Regulation in Luteinized Granulosa Cells Endocrinology, October 1, 2003; 144(10): 4597 - 4606. [Abstract] [Full Text] [PDF]

				A. E. Falender, R. Lanz, D. Malenfant, L. Belanger, and J. S. Richards Differential Expression of Steroidogenic Factor-1 and FTF/LRH-1 in the Rodent Ovary Endocrinology, August 1, 2003; 144(8): 3598 - 3610. [Abstract] [Full Text] [PDF]

				D. L. Russell, S. A. Ochsner, M. Hsieh, S. Mulders, and J. S. Richards Hormone-Regulated Expression and Localization of Versican in the Rodent Ovary Endocrinology, March 1, 2003; 144(3): 1020 - 1031. [Abstract] [Full Text] [PDF]

				V. Sriraman, S. C. Sharma, and J. S. Richards Transactivation of the Progesterone Receptor Gene in Granulosa Cells: Evidence that Sp1/Sp3 Binding Sites in the Proximal Promoter Play a Key Role in Luteinizing Hormone Inducibility Mol. Endocrinol., March 1, 2003; 17(3): 436 - 449. [Abstract] [Full Text] [PDF]

				T. Sekiguchi, T. Mizutani, K. Yamada, T. Yazawa, H. Kawata, M. Yoshino, T. Kajitani, T. Kameda, T. Minegishi, and K. Miyamoto Transcriptional Regulation of the Epiregulin Gene in the Rat Ovary Endocrinology, December 1, 2002; 143(12): 4718 - 4729. [Abstract] [Full Text] [PDF]

				J.J. Peluso, G. Fernandez, A. Pappalardo, and B.A. White Membrane-Initiated Events Account for Progesterone's Ability to Regulate Intracellular Free Calcium Levels and Inhibit Rat Granulosa Cell Mitosis Biol Reprod, August 1, 2002; 67(2): 379 - 385. [Abstract] [Full Text] [PDF]

				L. M. Salvador, Y. Park, J. Cottom, E. T. Maizels, J. C. R. Jones, R. V. Schillace, D. W. Carr, P. Cheung, C. D. Allis, J. L. Jameson, et al. Follicle-stimulating Hormone Stimulates Protein Kinase A-mediated Histone H3 Phosphorylation and Acetylation Leading to Select Gene Activation in Ovarian Granulosa Cells J. Biol. Chem., October 19, 2001; 276(43): 40146 - 40155. [Abstract] [Full Text] [PDF]

				C. Guo, L. Savage, K. D. Sarge, and O.-K. Park-Sarge Gonadotropins Decrease Estrogen Receptor-{beta} Messenger Ribonucleic Acid Stability in Rat Granulosa Cells Endocrinology, June 1, 2001; 142(6): 2230 - 2237. [Abstract] [Full Text] [PDF]

				J.-I. Park, W.-J. Kim, L. Wang, H.-J. Park, J. Lee, J.-H. Park, H.-B. Kwon, A. Tsafriri, and S.-Y. Chun Involvement of progesterone in gonadotrophin-induced pituitary adenylate cyclase-activating polypeptide gene expression in pre-ovulatory follicles of rat ovary Mol. Hum. Reprod., March 1, 2000; 6(3): 238 - 245. [Abstract] [Full Text] [PDF]

				B. L. Bohnsack, M. Szabo, S. M. Kilen, D. H.Y. Tam, and N. B. Schwartz Follistatin Suppresses Steroid-Enhanced Follicle-Stimulating Hormone Release In Vitro in Rats Biol Reprod, March 1, 2000; 62(3): 636 - 641. [Abstract] [Full Text]

				C. Ko, Y.-H. In, and O.-K. Park-Sarge Role of Progesterone Receptor Activation in Pituitary Adenylate Cyclase Activating Polypeptide Gene Expression in Rat Ovary Endocrinology, November 1, 1999; 140(11): 5185 - 5194. [Abstract] [Full Text]

				M. L. O’Brien, K. Park, Y. In, and O.-K. Park-Sarge Characterization of Estrogen Receptor-{beta} (ER{beta}) Messenger Ribonucleic Acid and Protein Expression in Rat Granulosa Cells Endocrinology, October 1, 1999; 140(10): 4530 - 4541. [Abstract] [Full Text]

				J. W. Clemens, R. L. Robker, W. L. Kraus, B. S. Katzenellenbogen, and J. S. Richards Hormone Induction of Progesterone Receptor (PR) Messenger Ribonucleic Acid and Activation of PR Promoter Regions in Ovarian Granulosa Cells: Evidence for a Role of Cyclic Adenosine 3',5'-Monophosphate but Not Estradiol Mol. Endocrinol., August 1, 1998; 12(8): 1201 - 1214. [Abstract] [Full Text]

				T. N. Alliston, A. C. Maiyar, P. Buse, G. L. Firestone, and J. S. Richards Follicle Stimulating Hormone-Regulated Expression of Serum/Glucocorticoid-Inducible Kinase in Rat Ovarian Granulosa Cells: A Functional Role for the Sp1 Family in Promoter Activity Mol. Endocrinol., December 1, 1997; 11(13): 1934 - 1949. [Abstract] [Full Text] [PDF]

				M. Byers, G. G. J. M. Kuiper, J.-A. Gustafsson, and O.-K. Park-Sarge Estrogen Receptor-{beta} mRNA Expression in Rat Ovary: Down-Regulation by Gonadotropins Mol. Endocrinol., February 1, 1997; 11(2): 172 - 182. [Abstract] [Full Text]