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Originally published In Press as doi:10.1074/jbc.M210801200 on January 27, 2003

J. Biol. Chem., Vol. 278, Issue 14, 12263-12270, April 4, 2003
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The Islet beta  Cell-enriched RIPE3b1/Maf Transcription Factor Regulates pdx-1 Expression*

Susan E. SamarasDagger , Li ZhaoDagger , Anna Means§, Eva HendersonDagger , Taka-aki MatsuokaDagger , and Roland SteinDagger

From the Dagger  Department of Molecular Physiology and Biophysics and the § Department of Surgical Oncology, Vanderbilt University Medical Center, Nashville, Tennessee 37232

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 mass (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 comprises the RIPE3b1 transcription factor. These results strongly suggest that RIPE3b1/Maf has an important role in generating and maintaining physiologically functional beta  cells.

* This work was supported by National Institutes of Health Grants RO1 DK50203 and P01 DK42502 (to R. S.) and Juvenile Diabetes Research Foundation Grants JDRF 398212 (to S. E. S.) and 32001678 (to T. M.). Partial support was provided by the Vanderbilt University Diabetes Research and Training Center, Molecular Biology Core Laboratory (Public Health Service Grant P60 DK20593).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 615-322-7026; Fax: 615-322-7236; E-mail: Roland.Stein@mcmail.vanderbilt.edu.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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