Abstract
Background/Aims: Molecular mechanisms underlying transcriptional rat multidrug-resistance protein 2 ( Mrp2, Abcc2) gene regulation are mostly unclear. Given the presence of putative binding sites for the Y-box binding protein YB-1 in the regulatory sequence, its trans-regulatory influence was analyzed. Methods: Reporter assays in HepG2 cells with various Mrp2 deletion constructs in the absence and presence of co-transfected YB-1 were performed. DNA binding studies with recombinant YB-1 protein and nuclear extracts obtained from HepG2 cells and rat liver tissue were carried out. Results: The minimal promoter sequence was confined to the proximal 186 bp. A YB-1 responsive element, Mrp2 YRE-1, was mapped at −186/−157, which exhibits specific YB-1 binding. YB-1 acts as a potent repressor of Mrp2 promoter activity in vitro. Conclusions: Constitutive Mrp2 gene expression is conferred through the proximal −186 bp. YB-1 acts as a repressor in vitro by specific binding to a defined element in the proximal promoter sequence.
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