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Volume 270, Number 43, Issue of October 27, 1995 pp. 25468-25474
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Identification and Characterization of a Hepatoma Cell-specific Enhancer in the Mouse Multidrug Resistance mdr1b Promoter

(Received for publication, June 6, 1995; and in revised form, August 16, 1995)

Renduo Song Masahidei Ikeguchi Ge Zhou M. Tien Kuo

The expression of multidrug resistance/P-glycoprotein genes mdr1b(mdr1) and mdr1a(mdr3) is elevated during hepatocarcinogenesis. To investigate the regulation of mdr1b gene expression, we used transient transfection expression assays of reporter constructs containing various 5`-mdr1b flanking sequences in hepatoma and non-hepatoma cells. We found that nucleotides -233 to -116 preferentially enhanced the expression of reporter gene in mouse hepatoma cell lines in an orientation- and promoter context-independent manner. DNase I footprinting using nuclear extracts prepared from hepatoma and non-hepatoma cells identified four protein binding sites at nucleotides -205 to -186 (site A), -181 to -164 (site B), -153 to -135 (site C), and -128 to -120 (site D). Further analyses revealed that, while site B alone played a major part for the enhancer function, sites A and B combined conferred full enhancer activity. Site-directed mutagenesis results also supported these results. Gel retardation experiments using oligonucleotide competitors revealed that the site B contains a dominant binding protein. This is the first report demonstrating a cell type-specific enhancer in the mdr locus. The role of this enhancer in the activation of mdr1b gene during hepatocarcinogenesis is discussed.




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G. Zhou and M. T. Kuo
Wild-type p53-mediated Induction of Rat mdr1b Expression by the Anticancer Drug Daunorubicin
J. Biol. Chem., June 19, 1998; 273(25): 15387 - 15394.
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Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.