title: Liganded pregnane X receptor represses the human sulfotransferase SULT1E1 promoter through disrupting its chromatin structure.
abstract: Pregnane X receptor (PXR), acting as a xenobiotic-activated transcription factor, regulates the hepatic metabolism of therapeutics as well as endobiotics such as steroid hormones. Given our finding that PXR activation by rifampicin (RIF) represses the estrogen sulfotransferase (SULT1E1) gene in human primary hepatocytes and hepatocellular carcinoma Huh7 cells, here we have investigated the molecular mechanism of this repression. First the PXR-responsive enhancer was delineated to a 100 bp sequence (-1000/-901), which contains three half sites that constitute the overlapping direct repeat 1 (DR1) and direct repeat 2 (DR2) motifs and two forkhead factor binding sites. siRNA knockdown, chromatin immunoprecipitation and chromatin conformation capture assays were employed to demonstrate that hepatocyte nuclear factor 4α (HNF4α) bound to the PXR-responsive enhancer, and activated the enhancer by looping its position close to the proximal promoter. Upon activation by RIF, PXR indirectly interacted with the enhancer, decreasing the interaction with HNF4α and dissolving the looped SULT1E1 promoter with deacetylation of histone 3. Removal of the DR sites from the enhancer hampers the ability of HNF4α to loop the promoter and that of PXR to repress the promoter activity. Thus, PXR represses human SULT1E1, possibly attenuating the inactivation of estrogen.
authors.affiliation: Pharmacogenetics Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709, USA.
authors.forename: Susumu
authors.initials: S
authors.lastname: Kodama
authors.forename: Fardin
authors.initials: F
authors.lastname: Hosseinpour
authors.forename: Joyce A
authors.initials: JA
authors.lastname: Goldstein
authors.forename: Masahiko
authors.initials: M
authors.lastname: Negishi
mesh_terms: D002478:Cells, Cultured; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D051542:Hepatocyte Nuclear Factor 3-beta / Q000378:metabolism; D051557:Hepatocyte Nuclear Factor 4 / Q000378:metabolism; D006801:Humans; D008024:Ligands; D000077297:Pregnane X Receptor; D011401:Promoter Regions, Genetic; D011987:Receptors, Steroid / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D015238:Sulfotransferases / Q000235:genetics*
chemical_list: D002843:Chromatin; D051557:Hepatocyte Nuclear Factor 4; D008024:Ligands; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid; D012097:Repressor Proteins; D051542:Hepatocyte Nuclear Factor 3-beta; D015238:Sulfotransferases; C020122:estrone sulfotransferase