Nucleic Acids Research, 1992, Vol. 20, No. 6 1393-1399
© 1992
MOLECULAR BIOLOGY |
Structure of the rat gonadotropin releasing hormone (rGnRH) gene promoter and functional analysis in hypothalamic cells
1Department of Medicine, University of Colorado Health Sciences Center Denver, CO 80220, USA 2Research Service, Veterans Affairs Medical Center Denver, CO 80220, USA
*To whom correspondence should be addressed at Section of Endocrinology (111H), VAMC, 1055 Clermont Street, Denver, CO 80220, USA
Received October 9, 1991. Revised February 21, 1992. Accepted February 21, 1992.
The gonadotropin releasing hormone (GnRH) gene encodes a protein which plays a critical role in mammalian reproductive physiology. Its expression is predominantly restricted to the hypothalamus although It has also been described In the placenta. To begin to determine the promoter elements important for tissue specific expression and to examine the mechanisms of developmental and hormonal regulation of the rat GnRH (rGnRH) gene, we cloned the rGnRH gene from a rat liver genomic DNA library. The nucleotide sequence of greater than 3 kb of 5'-flanking region was determined. The transcriptional initiation site in rat hypothalamic tissue and a mouse hypothalamic cell line were mapped by primer extension analysis and found to be different. In addition, transient transfection studies demonstrated that multiple regions of the distal promoter are Important for tissue specific and basal promoter activity in hypothalamic cells. Furthermore, in these cells a potent activation region resides between – 3026 and –1031 bp and suppressor region between –1031 and –903 bp upstream of the transcriptional start site. We conclude that different portions of the 5'-flanking region, which are activating and suppressing in nature, are critical for hypothalamic expression of the rGnRH gene.
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