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Papers In Press, published online ahead of print May 18, 2000
Department of Physiology and Pharmacology, Texas A&M University, College Station, TX 77843-4466
Corresponding Author: kramos{at}cvm.tamu.edu
Studies were conducted to evaluate the negative regulatory function of the rat (r)GST-Ya antioxidant/electrophile response element (ARE/EpRE) in vascular smooth muscle cells (vSMCs). C/EBP- was found to interact with ARE/EpRE in the rGST-Ya promoter, and that aryl hydrocarbon receptor (AhR) is present within the protein complex binding to the C/EBP site. Overexpression of C/EBP- or C/EBP- repressed, while AhR protein enhanced, 1.6CAT reporter activity in cells treated with 3 M benzo(a)pyrene (BaP). Overexpression of CREB binding protein (CBP) nullified ARE/EpRE-mediated repression of rGST-Ya transcription. Human adenovirus E1A protein abrogated co-transactivation by CBP, while an E1A mutant (E1A2/36) did not. Overexpression of C/EBPs abrogated stimulation of 1.6CAT by CBP or AhR alone, or in combination, irrespective of BaP treatment. Similar profiles were observed using an AhRECAT reporter construct. The C/EBP site within the ARE/EpRE inhibited chemical inducibility of the AhRE, the major BaP-responsive enhancer of 1.6CAT in vSMCs. We conclude that multiple mechanisms mediate negative regulation of rGST-Ya gene in vSMCs, most significant of which are that C/EBP-inhibits AhRE and ARE/EpRE inducibility of rGST-Ya in vSMCs, limiting CBP levels compromise induction of rGST-Ya gene and, AhR alone, or in combination with C/EBP- functions as a repressor of the ARE/EpRE.
J. Biol. Chem, 10.1074/jbc.M000405200
Submitted on January 19, 2000
Revised on April 12, 2000
Accepted on May 18, 2000
A CCAAT/enhancer binding protein (C/EBP) site within antioxidant/electrophile response element (ARE/EpRE) along with CREB binding protein (CBP) participate in the negative regulation of RatGST-Ya gene in vascular smooth muscle cells
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