title: Identification of pregnane X receptor binding sites in the regulatory regions of genes involved in bile acid homeostasis.
abstract: The nuclear receptor pregnane X receptor (PXR) acts as a sensor for a broad variety of natural and synthetic lipophilic compounds, such as bile acids and rifampicin, and regulates the expression of proteins that are involved in the metabolism and transport of these compounds. PXR binds as a heterodimer with the retinoid X receptor (RXR) to specific DNA sites, called response elements (REs), within the promoter regions of genes it activates transcriptionally. In this study we created a position weight matrix (PWM) for PXR-RXR heterodimers that took the relative in vitro binding strength and not only the sequence of natural and synthetic PXR binding sites (PXREs) into account. We further extended the discriminatory power of the matrix by including the variation of the dinucleotides 5'-flanking the hexameric binding motifs, which we show to have a significant effect on PXR binding ability. To test this PWM, it was used to screen the promoter regions of the human organic anion transport protein 2 (OATP2) and small heterodimer partner 1 (SHP1) genes. This resulted in the identification of 17 potential PXREs, of which seven bound PXR-RXR heterodimers in vitro. Furthermore, in HepG2 human hepatoma cells, PXR and RXR occupied chromatin regions that contained four of these REs. Induction of OATP2 and SHP1 mRNA expression by rifampicin confirmed that both genes are primary human PXR responding genes. This observation increases the understanding of the physiological role of PXR in the homeostasis of bile acids in humans.
authors.affiliation: Department of Biochemistry, University of Kuopio, FIN-70211 Kuopio, Finland.
authors.forename: Christian
authors.initials: C
authors.lastname: Frank
authors.forename: Harri
authors.initials: H
authors.lastname: Makkonen
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Dunlop
authors.forename: Merja
authors.initials: M
authors.lastname: Matilainen
authors.forename: Sami
authors.initials: S
authors.lastname: Väisänen
authors.forename: Carsten
authors.initials: C
authors.lastname: Carlberg
mesh_terms: D001647:Bile Acids and Salts / Q000378:metabolism*; D001665:Binding Sites; D045744:Cell Line, Tumor; D002843:Chromatin; D019281:Dimerization; D005786:Gene Expression Regulation / Q000187:drug effects; D006706:Homeostasis; D006801:Humans; D027381:Liver-Specific Organic Anion Transporter 1 / Q000235:genetics; D000077297:Pregnane X Receptor; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000502:physiology*; D011987:Receptors, Steroid / Q000502:physiology*; D020218:Response Elements / Q000502:physiology*; D012293:Rifampin / Q000494:pharmacology; D014162:Transfection
chemical_list: D001647:Bile Acids and Salts; D002843:Chromatin; D027381:Liver-Specific Organic Anion Transporter 1; D000077297:Pregnane X Receptor; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; C099713:nuclear receptor subfamily 0, group B, member 2; D012293:Rifampin