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Am J Physiol Renal Physiol (December 18, 2001). doi:10.1152/ajprenal.00268.2001
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Articles in PresS, published online ahead of print December 18, 2001
Am J Physiol Renal Physiol, 10.1152/ajprenal.00268.2001
Submitted on August 27, 2001
Accepted on November 23, 2001

GENOMIC ORGANIZATION OF THE 5' END OF HUMAN ßENAC AND PRELIMINARY CHARACTERIZATION OF ITS PROMOTER

Christie P Thomas1*, Randy W Loftus2, Kang Z Liu2, and Omar A Itani2

1 Internal Medicine, University of Iowa, Iowa City, IA, USA; Veterans Affairs Medical Center, Iowa City, IA, USA
2 Internal Medicine, University of Iowa, Iowa City, IA, USA

* To whom correspondence should be addressed. E-mail: christie-thomas{at}uiowa.edu.

The mRNA for the beta subunit of the epithelial sodium channel (ßENaC) is regulated developmentally and, in some tissues, in response to corticosteroids. To understand the mechanisms of transcriptional regulation of the human ßENaC gene we characterized the 5' end of the gene and its 5' flanking regions. Adaptor-ligated human kidney and lung cDNA were amplified by 5' RACE and transcription start sites of two 5' variant transcripts were determined by nuclease protection or primer extension assays. Cosmid clones that contain the 5' end of the gene were isolated and analysis of these clones indicated that alternate 1st exons ~1.5 kb apart and ~ 45 kb upstream of a common 2nd exon formed the basis of these transcripts. Genomic fragments that included the proximal 5' flanking region of either transcript were able to direct expression of a reporter gene in lung epithelia and to bind Sp1 in nuclear extracts confirming the presence of separate promoters that regulate ßENaC expression.




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