title: Multi-species analyses of direct activators of the constitutive androstane receptor.
abstract: The constitutive androstane receptor (CAR; NR1I3) is a member of the nuclear receptor superfamily and functions as an important xenochemical sensor and transcriptional modulator in mammalian cells. Upon chemical activation, CAR undergoes nuclear translocation and heterodimerization with the retinoid X receptor subsequent to its DNA target interaction. CAR is unusual among nuclear receptors in that it possesses a high level of constitutive activity in cell-based assays, obscuring the detection of ligand activators. However, a human splice variant of CAR, termed CAR3, exhibits negligible constitutive activity. In addition, CAR3 is activated by ligands with similar specificity as the reference form of the receptor. In this study, we hypothesized that similar CAR3 receptors could be constructed across various mammalian species' forms of CAR that would preserve species-specific ligand responses, thus enabling a more sensitive and differential screening assessment of CAR response among animal models. A battery of CAR3 receptors was produced in mouse, rat, and dog and comparatively evaluated with selected ligands together with human CAR1 and CAR3 in mammalian cell reporter assays. The results demonstrate that the 5-amino acid insertion that typifies human CAR3 also imparts ligand-activated receptor function in other species' CAR while maintaining signature responses in each species to select CAR ligands. These variant constructs permit in vitro evaluation of differential chemical effector responses across species and coupled with in vivo assays, the species-selective contributions of CAR in normal physiology and in disease processes such as hepatocarcinogenesis.
authors.affiliation: Center for Molecular Toxicology and Carcinogenesis, Department of Veterinary and Biomedical Sciences, Penn State University, University Park, Pennsylvania 16802, USA. cjo10@psu.edu
authors.forename: Curtis J
authors.initials: CJ
authors.lastname: Omiecinski
authors.forename: Denise M
authors.initials: DM
authors.lastname: Coslo
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Laurenzana
authors.forename: Richard C
authors.initials: RC
authors.lastname: Peffer
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002478:Cells, Cultured; D000090702:Constitutive Androstane Receptor; D004285:Dogs; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000737:chemistry / Q000187:drug effects; D006801:Humans; D017964:Itraconazole / Q000494:pharmacology; D008024:Ligands; D008099:Liver / Q000378:metabolism; D051379:Mice; D020033:Protein Isoforms; D022821:RNA Splice Sites; D012333:RNA, Messenger / Q000737:chemistry; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis* / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D000090702:Constitutive Androstane Receptor; D008024:Ligands; C000716327:NR1I3 protein, human; C000716328:Nr1i3 protein, mouse; C000716329:Nr1i3 protein, rat; D020033:Protein Isoforms; D022821:RNA Splice Sites; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D017964:Itraconazole