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Originally published In Press as doi:10.1074/jbc.M100563200 on April 24, 2001

J. Biol. Chem., Vol. 276, Issue 26, 23562-23571, June 29, 2001
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Pituitary Adenylate Cyclase-activating Polypeptide and Cyclic Adenosine 3',5'-Monophosphate Stimulate the Promoter Activity of the Rat Gonadotropin-releasing Hormone Receptor Gene via a Bipartite Response Element in Gonadotrope-derived Cells*

Hanna PincasDagger , Jean-Noël Laverrière, and Raymond Counis§

From the Endocrinologie Cellulaire et Moléculaire de la Reproduction, Université Pierre et Marie Curie, Centre National de la Recherche Scientifique, ESA 7080, 75252 Paris, France

Specific type I receptors for pituitary adenylate cyclase-activating polypeptide (PACAP) are present in gonadotrope cells of the anterior pituitary gland. By transient transfection of mouse gonadotrope-derived alpha T3-1 cells, which are direct targets for PACAP and express gonadotropin-releasing hormone receptor (GnRH-R), a marker of the gonadotrope lineage, we provide the first evidence that PACAP stimulates rat GnRH-R gene promoter activity. The EC50 of this stimulation is compatible with a mediation via activation of the cyclic AMP-dependent signaling pathway and, consistently, co-transfection of an expression vector expressing the protein kinase A inhibitor causes reduction in PACAP as well as cholera toxin-stimulated promoter activity. Deletion and mutational analyses indicate that PACAP activation necessitates a bipartite response element that consists of a first region (-272/-237) termed PACAP response element (PARE) I that includes a steroidogenic factor-1 (SF-1)-binding site and a second region (-136/-101) referred to as PARE II that contains an imperfect cyclic AMP response element. Gel shift experiments indicate the specific binding of the SF-1 and a potential SF-1-interacting factor to PARE I while a protein immunologically related to the cyclic AMP response element-binding protein interacts with PARE II. These findings suggest that PACAP might regulate the GnRH-R gene at the transcriptional level, providing novel insights into the regulation of pituitary-specific genes by hypothalamic hypophysiotropic signals.

* This work was supported by grants from the CNRS and the Université Pierre et Marie Curie.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Recipient of funds from the Ministère de l'Education Nationale de la Recherche et de la Technologie and from the Fondation pour la Recherche Médicale.

§ To whom correspondence should be addressed: Endocrinologie Cellulaire et Moléculaire de la Reproduction, Université Pierre & Marie Curie, CNRS ESA 7080, Case 244, 75252 Paris cedex 05, France. E-mail: Raymond.Counis@snv.jussieu.fr.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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