SpringerProtocols: Abstract: Transcriptional Regulation of UDP-Glucuronosyltransferases

Transcriptional Regulation of UDP-Glucuronosyltransferases

By: Anna Radominska-Pandya², Peter I. Mackenzie³, Wen Xie⁴

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Recent evidence has been provided that five nuclear receptors (NRs), (pregnane X receptor [PXR], constitutive androstane receptor [CAR], farnesoid X receptor [FXR] and peroxisome proliferator-activated receptor [PPARα and γ]) regulate the expression of UGT1A loci. This chapter presents an overview of the most recent developments in our understanding of transcriptional regulation of UDP-glucuronosyltransferase (UGT) genes, and specifically focuses on the regulation of UGTs via NRs, with a strong emphasis on the biochemical and molecular techniques applied to this area of investigation. Procedures are described for the isolation and analysis of UGT proximal and distal promoters. The use of transgenic animals in the study of the mechanism of regulation of UGT expression is described, as is the assessment of the effects of various gene inducers on expression of UGTs in HepG2 and Caco-2 cell culture. Techniques for transient transfection of HepG2 and Caco-2 cells with NRs and UGT promoter constructs are presented. In addition, general methods are provided for various molecular and biochemical methods used in the isolation and characterization of UGT promoters.

Affiliation(s): (2) Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR
(3) Department of Clinical Pharmacology, Flinders University, Bedford Park, Australia
(4) Center for Pharmacogenetics and Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA

Book Title: Drug Metabolism and Transport: Molecular Methods and Mechanisms

Series: Methods in Pharmacology and Toxicology | Pub. Date: Sep-24-2004 | Page Range: 133-171 | DOI: 10.1385/1-59259-832-3:133

Subject: Pharmacology/Toxicology

Key Words: Aryl hydrocarbon receptor - bile acids - bilirubin - (bio)flavonoids - CaCo-2 cells - constitutive androgen receptor - cytochrome P450 - DNase 1 footprinting - electrophoretic mobility shift assay - glucuronidation assays - HepG2 cells - luciferase reporter constructs - mouse: “humanized” - mouse: transgenic - Northern blot hybridization - peroxisome proliferator-activated receptor - pregnane X receptor - retinoids - reverse transcriptase-polymerase chain reaction - steroid hormones - transfection: transient - UDP-glucuronosyltransferase - UGT promoter constructs - UGT transcriptional regulation - xenobiotic response element

References

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