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Am J Physiol Gastrointest Liver Physiol (February 5, 2003). doi:10.1152/ajpgi.00483.2002
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Submitted on November 7, 2002
Accepted on February 4, 2003

High-level activation by a duodenum specific enhancer requires functional GATA binding sites

Mary R. Dusing1, Elizabeth A. Florence1, and Dan A. Wiginton1*

1 Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA; Division of Developmental Biology, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH, USA

* To whom correspondence should be addressed. E-mail: dan.wiginton{at}chmcc.org.

The purine metabolic gene adenosine deaminase (ADA) is expressed at high levels in a well defined spatiotemporal pattern in the villous epithelium of proximal small intestine. A duodenum-specific enhancer module responsible for this expression pattern has been identified in the second intron of the human ADA gene. It has previously been shown that binding of the factor PDX-1 is essential for function of this enhancer. The studies presented here examine the proposed roles of GATA factors in the enhancer. Site-directed mutagenesis of the enhancer's GATA binding sites crippled enhancer function in ten lines of transgenic mice, with nine of the lines demonstrating less than 1% of normal activity. Detailed studies along the longitudinal axis of mouse small intestine indicate that GATA-4 and GATA-5 mRNA levels display a reciprocal pattern, with low levels of GATA-6 throughout. Interestingly, gel shift studies with duodenal nuclear extracts showed binding only by GATA-4.




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