2015
DOI: 10.4236/abc.2015.52008
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Identification of CAR/RXR<i>α</i> Hetero-dimer Binding Sites in the Human Genome by a Modified Yeast One-Hybrid Assay

Abstract: The constitutive androstane receptor (CAR) is a transcription factor that belongs to the nuclear receptor superfamily. CAR binds as a heterodimer with the retinoid X receptor α (RXRα) to CAR response elements (CAREs) and regulates the expression of various drug metabolizing enzymes and transporters. To identify CAR/RXRα binding sites in the human genome, we performed a modified yeast one-hybrid assay that enables rapid and efficient identification of genomic targets for DNA-binding proteins. DNA fragments were… Show more

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Cited by 5 publications
(3 citation statements)
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References 44 publications
(51 reference statements)
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“…Nevertheless, genome-wide chromatin immunoprecipitation (ChIP) and deep sequencing of immunoprecipitated DNAs (ChIP-Seq) identified DR4 as the most frequent PXR- associated recruitment sites in mouse liver [30]. DR4 in the human genome is a preferred DNA-binding site for the CAR/RXR heterodimer as well, as recently observed in a modified yeast one- hybrid assay [31]. DR3 is the prevalent VDR-binding site at genomic regions that contain primary VDR target genes.…”
Section: Nuclear Receptors (Nrs) Response Elements Gene Regulatiomentioning
confidence: 98%
“…Nevertheless, genome-wide chromatin immunoprecipitation (ChIP) and deep sequencing of immunoprecipitated DNAs (ChIP-Seq) identified DR4 as the most frequent PXR- associated recruitment sites in mouse liver [30]. DR4 in the human genome is a preferred DNA-binding site for the CAR/RXR heterodimer as well, as recently observed in a modified yeast one- hybrid assay [31]. DR3 is the prevalent VDR-binding site at genomic regions that contain primary VDR target genes.…”
Section: Nuclear Receptors (Nrs) Response Elements Gene Regulatiomentioning
confidence: 98%
“…Using experimentally-validated sequences for the CAR response elements (CREs) [ 21 ], the promoter sequences obtained (as above) were examined for the presence of CREs using Snapgene gene viewer software.…”
Section: Methodsmentioning
confidence: 99%
“…Likewise, point mutations within the androgen receptor (AR) LBD – associated with androgen insensitivity syndromes – decrease the DNA-binding affinity of the AR homodimer, while leaving intact the ligand-binding affinity (Helsen, et al, 2012). Additionally, distinct ligand-dependent recognition patterns of promoter DNA by CAR have been identified (Cherian, Lin, Wu, & Chen, 2015; Hosoda, et al, 2015; Li, et al, 2015; H. Wang, et al, 2003).…”
Section: Linking Ligand and Dna Through Allosterymentioning
confidence: 99%