Endocrinology, Vol 135, 702-711, Copyright © 1994 by Endocrine Society

ARTICLES

Two messenger ribonucleic acids encoding the common beta B-chain of inhibin and activin have distinct 5'-initiation sites and are differentially regulated in rat granulosa cells

JC Dykema and KE Mayo
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208.

Inhibin and activin gene expression in the ovary and in cultured granulosa cells is modulated by the pituitary gonadotropins FSH and LH. In granulosa cells, two messenger RNAs (mRNAs) of 4.3 and 3.3 kilobases (kb) encoding the common beta B-chain of inhibin and activin are observed. We demonstrate here that FSH or pharmacological agents that elevate intracellular levels of cAMP induce expression of the 4.3-kb mRNA to a greater extent than that of the 3.3-kb mRNA. To elucidate the mechanism(s) by which the two beta B transcripts are generated, we characterized the rat beta B gene and used probes from various regions of the gene to examine the structures of the beta B mRNAs. RNA blot analysis using probes from the 5'-nontranslated and flanking regions of the beta B gene revealed that the 4.3- and 3.3-kb transcripts differ in the length of their 5'-nontranslated regions. S1 nuclease protection and primer extension assays were used to map two adjacent transcriptional start sites 1051 and 1052 basepairs (bp) up-stream of the most 3'-start site previously reported for the beta B gene. These novel start sites are used to generate the 4.3-kb mRNA, whereas transcription of the 3.3-kb mRNA initiates at down-stream start sites. DNA sequence analysis did not reveal any consensus CCAAT or TATA box elements up-stream of the novel start sites. Multiple potential binding sites for the transcription factors SP1 and activator protein-2 are present throughout a region of the beta B gene from -1461 to 82 bp. Fusion genes were constructed that contain beta B sequences from -1460 to 14 bp, -1460 to -914 bp, and -913 to 14 bp controlling luciferase reporter gene expression. Analysis of beta B promoter activity in transiently transfected primary granulosa cells demonstrated that sequences flanking both the up- and down-stream transcriptional start sites have substantial basal promoter activity; however, forskolin did not affect expression of the beta B/luciferase fusion genes.

This article has been cited by other articles:

				J. L. Kipp, S. M. Kilen, T. K. Woodruff, and K. E. Mayo Activin Regulates Estrogen Receptor Gene Expression in the Mouse Ovary J. Biol. Chem., December 14, 2007; 282(50): 36755 - 36765. [Abstract] [Full Text] [PDF]

				J. L. Kipp, S. M. Kilen, S. Bristol-Gould, T. K. Woodruff, and K. E. Mayo Neonatal Exposure to Estrogens Suppresses Activin Expression and Signaling in the Mouse Ovary Endocrinology, May 1, 2007; 148(5): 1968 - 1976. [Abstract] [Full Text] [PDF]

				S. E. Harris, A. L. Chand, I. M. Winship, K. Gersak, Y. Nishi, T. Yanase, H. Nawata, and A. N. Shelling INHA promoter polymorphisms are associated with premature ovarian failure Mol. Hum. Reprod., November 1, 2005; 11(11): 779 - 784. [Abstract] [Full Text] [PDF]

				Y Okuma, A E O'Connor, J A Muir, P G Stanton, D M de Kretser, and M P Hedger Regulation of activin A and inhibin B secretion by inflammatory mediators in adult rat Sertoli cell cultures J. Endocrinol., October 1, 2005; 187(1): 125 - 134. [Abstract] [Full Text] [PDF]

				H. Alam, E. T. Maizels, Y. Park, S. Ghaey, Z. J. Feiger, N. S. Chandel, and M. Hunzicker-Dunn Follicle-stimulating Hormone Activation of Hypoxia-inducible Factor-1 by the Phosphatidylinositol 3-Kinase/AKT/Ras Homolog Enriched in Brain (Rheb)/Mammalian Target of Rapamycin (mTOR) Pathway Is Necessary for Induction of Select Protein Markers of Follicular Differentiation J. Biol. Chem., May 7, 2004; 279(19): 19431 - 19440. [Abstract] [Full Text] [PDF]

				S. Ramaswamy, G. R. Marshall, C. R. Pohl, R. L. Friedman, and T. M. Plant Inhibitory and Stimulatory Regulation of Testicular Inhibin B Secretion by Luteinizing Hormone and Follicle-Stimulating Hormone, Respectively, in the Rhesus Monkey (Macaca mulatta) Endocrinology, April 1, 2003; 144(4): 1175 - 1185. [Abstract] [Full Text] [PDF]

				Y. Wang and W. Ge Involvement of Cyclic Adenosine 3',5'-Monophosphate in the Differential Regulation of Activin {beta}A and {beta}B Expression by Gonadotropin in the Zebrafish Ovarian Follicle Cells Endocrinology, February 1, 2003; 144(2): 491 - 499. [Abstract] [Full Text] [PDF]

				J.-S. Roh, J. Bondestam, S. Mazerbourg, N. Kaivo-Oja, N. Groome, O. Ritvos, and A. J. W. Hsueh Growth Differentiation Factor-9 Stimulates Inhibin Production and Activates Smad2 in Cultured Rat Granulosa Cells Endocrinology, January 1, 2003; 144(1): 172 - 178. [Abstract] [Full Text] [PDF]

				C. Marchetti, M. Hamdane, V. Mitchell, K. Mayo, L. Devisme, J.M. Rigot, J.C. Beauvillain, E. Hermand, and A. Defossez Immunolocalization of Inhibin and Activin {alpha} and {beta}B Subunits and Expression of Corresponding Messenger RNAs in the Human Adult Testis Biol Reprod, January 1, 2003; 68(1): 230 - 235. [Abstract] [Full Text] [PDF]

				C. Welt, Y. Sidis, H. Keutmann, and A. Schneyer Activins, Inhibins, and Follistatins: From Endocrinology to Signaling. A Paradigm for the New Millennium Experimental Biology and Medicine, October 1, 2002; 227(9): 724 - 752. [Abstract] [Full Text] [PDF]

				K. Y. Arai, K.-i. Ohshima, G. Watanabe, K. Arai, K. Uehara, and K. Taya Dynamics of Messenger RNAs Encoding Inhibin/Activin Subunits and Follistatin in the Ovary During the Rat Estrous Cycle Biol Reprod, April 1, 2002; 66(4): 1119 - 1126. [Abstract] [Full Text] [PDF]

				J. Liu, C. Hyden-Granskog, and R. Voutilainen Gonadotrophins inhibit and activin induces expression of inhibin/activin {beta}B subunit mRNA in cultured human granulosa-luteal cells Mol. Hum. Reprod., April 1, 2001; 7(4): 319 - 323. [Abstract] [Full Text] [PDF]

				C. K. Welt and A. L. Schneyer Differential Regulation of Inhibin B and Inhibin A by Follicle-Stimulating Hormone and Local Growth Factors in Human Granulosa Cells from Small Antral Follicles J. Clin. Endocrinol. Metab., January 1, 2001; 86(1): 330 - 336. [Abstract] [Full Text]

				Z.-M. Feng, A. Z. Wu, Z. Zhang, and C.-L. C. Chen GATA-1 and GATA-4 Transactivate Inhibin/Activin {beta}-B-Subunit Gene Transcription in Testicular Cells Mol. Endocrinol., November 1, 2000; 14(11): 1820 - 1835. [Abstract] [Full Text]

				D. J. Hecht, A. J. Davis, C. F. Brooks, and P. A. Johnson Molecular Cloning and Expression Analysis of the Complementary Deoxyribonucleic Acid for Chicken Inhibin/Activin {beta}B Subunit Biol Reprod, May 1, 2000; 62(5): 1128 - 1134. [Abstract] [Full Text]

				S. Ramaswamy, G. R. Marshall, A. S. McNeilly, and T. M. Plant Dynamics of the Follicle-Stimulating Hormone (FSH)-Inhibin B Feedback Loop and Its Role in Regulating Spermatogenesis in the Adult Male Rhesus Monkey (Macaca mulatta) as Revealed by Unilateral Orchidectomy Endocrinology, January 1, 2000; 141(1): 18 - 27. [Abstract] [Full Text] [PDF]

				A. M. Ardekani, J. C. D. Romanelli, and K. E. Mayo Structure of the Rat Inhibin and Activin {beta}A-Subunit Gene and Regulation in an Ovarian Granulosa Cell Line Endocrinology, July 1, 1998; 139(7): 3271 - 3279. [Abstract] [Full Text] [PDF]

				Z.-M. Feng, A. Z. Wu, and C.-L. C. Chen Testicular GATA-1 Factor Up-Regulates the Promoter Activity of Rat Inhibin {alpha}-Subunit Gene in MA-10 Leydig Tumor Cells Mol. Endocrinol., March 1, 1998; 12(3): 378 - 390. [Abstract] [Full Text]

				K. Tanimoto, E. Yoshida, S. Mita, Y. Nibu, K. Murakami, and A. Fukamizu Human Activin beta A Gene. IDENTIFICATION OF NOVEL 5prime EXON, FUNCTIONAL PROMOTER, AND ENHANCERS J. Biol. Chem., December 20, 1996; 271(51): 32760 - 32769. [Abstract] [Full Text] [PDF]

				Y.-H. Lin, E. J. Shin, M. J. Campbell, and J. E. Niederhuber Transcription of the blk Gene in Human B Lymphocytes Is Controlled by Two Promoters J. Biol. Chem., October 27, 1995; 270(43): 25968 - 25975. [Abstract] [Full Text] [PDF]