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A more recent version of this article appeared on March 28, 2003
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M210801200v1
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Papers In Press, published online ahead of print January 27, 2003
J. Biol. Chem, 10.1074/jbc.M210801200
Submitted on October 22, 2002
Revised on January 21, 2003
Accepted on January 27, 2003

The islet beta cell-enriched RIPE3b1/Maf transcription factor regulates pdx-1 expression

Susan E. Samaras, Li Zhao, Anna Means, Eva Henderson, Taka-aki Matsuoka, and Roland Stein

Department of Molecular Physiology, Vanderbilt University, Nashville, TN 37232

Corresponding Author: roland.stein{at}vanderbilt.edu

Pancreatic duodenal homeobox factor–1, PDX-1, is required for pancreas development, islet cell differentiation, and the maintenance of beta cell function. Selective expression in the pancreas appears to be principally regulated by Area II, one of four conserved regulatory sequence domains found within the 5’-flanking region of the pdx-1 gene. Detailed mutagenesis studies have identified potential sites of interaction for both positive- and negative-acting factors within the conserved sequence blocks of Area II. The islet beta cell-enriched RIPE3b1 transcription factor, the activator of insulin C1 element-driven expression, was shown here to also stimulate Area II by binding to sequence blocks 4 and 5 (termed B4/5). Accordingly, B4/5 DNA-binding protein’s molecular weight (i.e. 46 kDa), binding specificity, and islet beta cell-enriched distribution were identical to RIPE3b1. Area II-mediated activation was also unaffected upon replacing B4/5 with the insulin C1/RIPE3b1 binding site. In addition, the chromatin immunoprecipitation assay showed that the Area II region of the endogenous pdx-1 gene was precipitated by an antiserum that recognizes the large Maf protein that composes the RIPE3b1 transcription factor. These results strongly suggest that RIPE3b1/Maf has an important role in generating and maintaining physiologically functional beta cells.


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