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Molecular Human Reproduction, Vol. 6, No. 8, 671-676, August 2000
© 2000 European Society of Human Reproduction and Embryology

Endocrinology

Potential regulation of GnRH gene by a steroidogenic factor-1-like protein

D.R. Corley1,2, X. Li1, Z.M. Lei1 and Ch.V. Rao1,3

1 Division of Basic Science Research, Department of Obstetrics and Gynecology, University of Louisville Health Sciences Center, Louisville, Kentucky 40292, USA

Abstract

Steroidogenic factor-1 (SF-1) is a member of an orphan nuclear hormone receptor superfamily. It plays a critical role in the development and function of the hypothalamic–pituitary–gonadal and adrenal axis. However, whether SF-1 can regulate transcription of gonadotrophin-releasing hormone (GnRH) gene is not known. To examine this possibility, we first over-expressed SF-1 and found that it not only decreased steady state GnRH messenger ribonucleic acid (mRNA) levels but also reduced its promoter activity in GT1-7 neurons. The inhibitory effect of SF-1 was lost when the 5'-flanking region of GnRH gene containing two distal (-1479 to –1474 bp and –1059 to –1054 bp) hexamers was deleted. Gel mobility shift assays showed that GT1-7 cell nuclear extracts contained a protein that formed a specific complex with synthetic oligonucleotides containing the two distal hexamers or a consensus SF-1 binding sequence. The migration of this complex was, however, slower than the complex formed with MA-10 cell nuclear extracts which were shown to contain a 53 kDa SF-1 protein. The addition of anti-SF-1 antibody supershifted the complex formed with MA-10, but not with GT1-7 cell nuclear extracts. The same antibody, however, detected a 60 kDa protein and immunostained nuclei of GT1-7 neurons. These results are consistent with GT1-7 neurons containing an SF-1-like protein that can bind to the distal hexamer sequences in the 5'-flanking region of rat GnRH gene to inhibit its transcription.

gene transcription/GnRH/GT1-7 neurons/MA-10 cells/SF-1

Notes

2 Present address: 14822 W. 71st Terrace, Shawnee, KS 66216, USA

3 To whom correspondence should be addressed at: Department of Ob/Gyn, 438 MDR Building, University of Louisville, Health Sciences Center, Louisville, KY 40292, USA. E-mail: cvrao001{at}gwise.louisville.edu


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