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Am J Physiol Renal Physiol (February 3, 2004). doi:10.1152/ajprenal.00319.2003
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Submitted on September 4, 2003
Accepted on January 23, 2004

Identification of A Novel Region in the Proximal Promoter of the Mouse Renin Gene Critical for Expression

Li Pan1, Craig A. Jones1, Sean T. Glenn1, and Kenneth W. Gross1*

1 Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, NY, USA

* To whom correspondence should be addressed. E-mail: gross{at}acsu.buffalo.edu.

An enhancer at -2.6 kb and a HOX - PBX-binding site at -60 bp have been demonstrated to be critical to expression of the mouse renin gene (Ren-1c) in As4.1 cells. In this report we show that a region (-197 to -70) immediately 5' to the HOX - PBX-binding site is also critical for Ren-1c expression. Deletion of this region in a construct containing 4.1 kb of Ren-1c 5' flanking sequence resulted in a 99% reduction in Ren-1c promoter activity in As4.1 cells, suggesting the pivotal role for the region in regulation of mouse renin gene. Electrophoretic mobility shift and supershift assays have identified two nuclear factor I (NFI)-binding sites and an Sp1/Sp3-binding site within the distal portion of the region (-197 to -103). Mutation of these three sites caused a 90% decrease in Ren-1c promoter activity. Mutational analysis and electrophoretic mobility shift assays have also identified three additional transcription factor-binding sites within the region from -103 to -69, each of which contributes to high-level expression of Ren-1c in As4.1 cells. Finally, we have shown that the Ren-1c enhancer is the target for endothelin-1 (ET-1)-induced inhibition of Ren-1c expression and the transcription factor-binding sites in the proximal promoter are required for the maximal ET-1 inhibitory effect.




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