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Endocrinology Vol. 138, No. 5 2098-2108
Copyright © 1997 by The Endocrine Society

Articles

Isolation and Characterization of the Rat Corticotropin-Releasing Hormone (CRH)-Binding Protein Gene: Transcriptional Regulation by Cyclic Adenosine Monophosphate and CRH1

Daniel N. Cortright, Ki A. Goosens, J. Shonee Lesh and Audrey F. Seasholtz

Department of Biological Chemistry (D.N.C., A.F.S.) and the Mental Health Research Institute (K.A.G., J.S.L., A.F.S.), The University of Michigan, Ann Arbor, Michigan 48109-0720

Address all correspondence and requests for reprints to: Audrey F. Seasholtz, Ph.D., Mental Health Research Institute, 205 Zina Pitcher Place, Ann Arbor, Michigan 48109-0720. E-mail: aseashol{at}umich.edu

The CRH-binding protein (CRH-BP) antagonizes the ACTH-releasing activity of the neuropeptide CRH in vitro. However, the function of CRH-BP in vivo and the molecular mechanisms that regulate CRH-BP expression are not well understood. In this study, the rat CRH-BP gene was characterized, and CRH-BP promoter sequences were identified. The rat CRH-BP gene spans almost 12 kilobases and contains 7 exons. Ribonuclease protection experiments indicate that transcription of the CRH-BP gene initiates at multiple sites in rat cerebral cortex. Transfection experiments with CRH-BP-reporter constructs, containing 88–3500 bp 5' flanking and 66 bp 5' untranslated DNA from the rat CRH-BP gene, demonstrate basal promoter activity in multiple cell lines. CRH-BP-reporter constructs also demonstrate positive regulation of promoter activity by cAMP in a variety of cell lines and by CRH in cells expressing the CRH receptor. The DNA sequences between -341 and -88 bp, including the cAMP response element-like sequence at -127 bp, are required for maximal cAMP and CRH regulation of CRH-BP promoter activity. These studies suggest that CRH-BP transcription in vivo may be positively regulated by cAMP and CRH.




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Copyright © 1997 by The Endocrine Society