This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager NURSA Molecule Pages Link Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sriraman, V. Articles by Richards, J. S. Search for Related Content PubMed PubMed Citation Articles by Sriraman, V. Articles by Richards, J. S.

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

Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030

Address all correspondence and requests for reprints to: Dr. JoAnne S. Richards, Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030. E-mail: joanner{at}bcm.tmc.edu.

LH induction of the progesterone receptor (PR) in granulosa cells is a central event in ovulation. To identify critical regions of the mouse PR promoter that confer LH inducibility in granulosa cells, a mouse PR promoter (-384/+680) genomic fragment was ligated to a luciferase reporter construct and transfected into primary cultures of granulosa cells. Forskolin/phorbol myristate (PMA) induced PR promoter-luciferase reporter activity in granulosa cells greater than 15-fold. A deletion construct comprised only of the distal promoter alone (-348/+64) was inactive. Conversely, deletion constructs eliminating putative distal promoter-regulatory elements that bind Sp1, nuclear factor Y, and GATA-4 as well as the transcription start site (+1) failed to reduce forskolin/PMA activation of reporter activity. Additional 5'-deletions identified a minimal promoter region (+420/+680) sufficient to bestow cAMP responsiveness approximately 8- to 10-fold. Two GC-rich regions Sp1(A)[+440/+461] and Sp1(B) [+473/+490] bound Sp1/Sp3. Site-directed mutagenesis of Sp1(A) and Sp1(B) reduced activity of the proximal (+357/+680) promoter reporter construct approximately 50% and 99%, respectively. When the same Sp1(B) mutation was introduced into the intact promoter (-145/+680), forskolin/PMA induction of promoter activity was reduced by 70–80%. When the distal GC box as well as the proximal Sp1(B) site were both mutated in the context of the intact promoter, inducibility of the transgene was even more severely reduced. The importance of these Sp1/Sp3 binding regions was confirmed in human MCF-7 cells and Drosophila SL2 cells. Collectively, these results indicate that the Sp1/Sp3 binding sites within the mouse PR proximal promoter are essential for transactivation of the gene by agonists in granulosa cells. The molecular mechanisms by which LH activates Sp1/Sp3 at this region within the PR gene remain unknown but do not involve changes in the binding of Sp1/Sp3 to the critical GC boxes. Rather, Sp1/Sp3 appear to recruit other factors to the promoter.

NURSA Molecule Pages Link:

Nuclear Receptors:   PR

Ligands:   17β-Estradiol

This article has been cited by other articles:

				V. Sriraman, U. Eichenlaub-Ritter, J. W. Bartsch, A. Rittger, S. M. Mulders, and J. S. Richards Regulated Expression of ADAM8 (a Disintegrin and Metalloprotease Domain 8) in the Mouse Ovary: Evidence for a Regulatory Role of Luteinizing Hormone, Progesterone Receptor, and Epidermal Growth Factor-Like Growth Factors Biol Reprod, June 1, 2008; 78(6): 1038 - 1048. [Abstract] [Full Text] [PDF]

				J. Kim, M. Sato, Q. Li, J. P. Lydon, F. J. DeMayo, I. C. Bagchi, and M. K. Bagchi Peroxisome Proliferator-Activated Receptor {gamma} Is a Target of Progesterone Regulation in the Preovulatory Follicles and Controls Ovulation in Mice Mol. Cell. Biol., March 1, 2008; 28(5): 1770 - 1782. [Abstract] [Full Text] [PDF]

				Y. Yamashita, I. Kawashima, Y. Yanai, M. Nishibori, J. S. Richards, and M. Shimada Hormone-Induced Expression of Tumor Necrosis Factor {alpha}-Converting Enzyme/A Disintegrin and Metalloprotease-17 Impacts Porcine Cumulus Cell Oocyte Complex Expansion and Meiotic Maturation via Ligand Activation of the Epidermal Growth Factor Receptor Endocrinology, December 1, 2007; 148(12): 6164 - 6175. [Abstract] [Full Text] [PDF]

				M. Shimada, Y. Yanai, T. Okazaki, Y. Yamashita, V. Sriraman, M. C. Wilson, and J. S. Richards Synaptosomal-Associated Protein 25 Gene Expression Is Hormonally Regulated during Ovulation and Is Involved in Cytokine/Chemokine Exocytosis from Granulosa Cells Mol. Endocrinol., October 1, 2007; 21(10): 2487 - 2502. [Abstract] [Full Text] [PDF]

				M. D Rudd, I. Gonzalez-Robayna, I. Hernandez-Gonzalez, N. L Weigel, W. E Bingman III, and J. S Richards Constitutively active FOXO1a and a DNA-binding domain mutant exhibit distinct co-regulatory functions to enhance progesterone receptor A activity J. Mol. Endocrinol., June 1, 2007; 38(6): 673 - 690. [Abstract] [Full Text] [PDF]

				D. L. Russell and R. L. Robker Molecular mechanisms of ovulation: co-ordination through the cumulus complex Hum. Reprod. Update, May 1, 2007; 13(3): 289 - 312. [Abstract] [Full Text] [PDF]

				N. Sher, N. Yivgi-Ohana, and J. Orly Transcriptional Regulation of the Cholesterol Side Chain Cleavage Cytochrome P450 Gene (CYP11A1) Revisited: Binding of GATA, Cyclic Adenosine 3',5'-Monophosphate Response Element-Binding Protein and Activating Protein (AP)-1 Proteins to a Distal Novel Cluster of cis-Regulatory Elements Potentiates AP-2 and Steroidogenic Factor-1-Dependent Gene Expression in the Rodent Placenta and Ovary Mol. Endocrinol., April 1, 2007; 21(4): 948 - 962. [Abstract] [Full Text] [PDF]

				C. Stocco, C. Telleria, and G. Gibori The Molecular Control of Corpus Luteum Formation, Function, and Regression Endocr. Rev., February 1, 2007; 28(1): 117 - 149. [Abstract] [Full Text] [PDF]

				M. Shimada, I. Hernandez-Gonzalez, I. Gonzalez-Robayna, and J. S. Richards Paracrine and Autocrine Regulation of Epidermal Growth Factor-Like Factors in Cumulus Oocyte Complexes and Granulosa Cells: Key Roles for Prostaglandin Synthase 2 and Progesterone Receptor Mol. Endocrinol., June 1, 2006; 20(6): 1352 - 1365. [Abstract] [Full Text] [PDF]

				V. Sriraman, M. D. Rudd, S. M. Lohmann, S. M. Mulders, and J. S. Richards Cyclic Guanosine 5'-Monophosphate-Dependent Protein Kinase II Is Induced by Luteinizing Hormone and Progesterone Receptor-Dependent Mechanisms in Granulosa Cells and Cumulus Oocyte Complexes of Ovulating Follicles Mol. Endocrinol., February 1, 2006; 20(2): 348 - 361. [Abstract] [Full Text] [PDF]

				P. Hanifi-Moghaddam, S. C. J. P. Gielen, H. J. Kloosterboer, M. E. De Gooyer, A. M. Sijbers, A. J. van Gool, M. Smid, M. Moorhouse, F. H. van Wijk, C. W. Burger, et al. Molecular Portrait of the Progestagenic and Estrogenic Actions of Tibolone: Behavior of Cellular Networks in Response to Tibolone J. Clin. Endocrinol. Metab., February 1, 2005; 90(2): 973 - 983. [Abstract] [Full Text] [PDF]

				K. M. H. Doyle, D. L. Russell, V. Sriraman, and J. S. Richards Coordinate Transcription of the ADAMTS-1 Gene by Luteinizing Hormone and Progesterone Receptor Mol. Endocrinol., October 1, 2004; 18(10): 2463 - 2478. [Abstract] [Full Text] [PDF]

				N. Gava, C. L. Clarke, K. Byth, R. L. Arnett-Mansfield, and A. deFazio Expression of Progesterone Receptors A and B in the Mouse Ovary during the Estrous Cycle Endocrinology, July 1, 2004; 145(7): 3487 - 3494. [Abstract] [Full Text] [PDF]

				S. J. Hirvonen-Santti, V. Sriraman, M. Anttonen, S. Savolainen, J. J. Palvimo, M. Heikinheimo, J. S. Richards, and O. A. Janne Small Nuclear RING Finger Protein Expression during Gonad Development: Regulation by Gonadotropins and Estrogen in the Postnatal Ovary Endocrinology, May 1, 2004; 145(5): 2433 - 2444. [Abstract] [Full Text] [PDF]

				Y.-H. Cheng, B. D. Richardson, M. A. Hubert, and S. Handwerger Isolation and Characterization of the Human Syncytin Gene Promoter Biol Reprod, March 1, 2004; 70(3): 694 - 701. [Abstract] [Full Text] [PDF]

				V. Sriraman and J. S. Richards Cathepsin L Gene Expression and Promoter Activation in Rodent Granulosa Cells Endocrinology, February 1, 2004; 145(2): 582 - 591. [Abstract] [Full Text] [PDF]