TY - JOUR T1 - The Major Human Pregnane X Receptor (PXR) Splice Variant, PXR.2, Exhibits Significantly Diminished Ligand-Activated Transcriptional Regulation JF - Drug Metabolism and Disposition JO - Drug Metab Dispos SP - 1295 LP - 1304 DO - 10.1124/dmd.108.025213 VL - 37 IS - 6 AU - Yvonne S. Lin AU - Kazuto Yasuda AU - Mahfoud Assem AU - Cynthia Cline AU - Joe Barber AU - Chia-Wei Li AU - Vladyslav Kholodovych AU - Ni Ai AU - J. Don Chen AU - William J. Welsh AU - Sean Ekins AU - Erin G. Schuetz Y1 - 2009/06/01 UR - http://dmd.aspetjournals.org/content/37/6/1295.abstract N2 - The pregnane X receptor (PXR; PXR.1) can be activated by structurally diverse lipophilic ligands. PXR.2, an alternatively spliced form of PXR, lacks 111 nucleotides encoding 37 amino acids in the ligand binding domain. PXR.2 bound a classic CYP3A4 PXR response element (PXRE) in electrophoretic mobility shift assays, but transfected PXR.2 failed to transactivate a CYP3A4-promoter-luciferase reporter plasmid in HepG2 cells treated with various PXR ligands. Cotransfection experiments showed that PXR.2 behaved as a dominant negative, interfering with PXR.1/rifampin activation of CYP3A4-PXRE-LUC. In HepG2 and LS180 cells stably transduced with PXR.1, PXR target genes (CYP3A4, MDR1, CYP2B6, and UGT1A1) were higher than mock-transduced cells in the absence of ligand and were further induced in the presence of rifampin. In contrast, PXR.2 stably introduced into the same host cells failed to induce target genes over levels in mock-transfected cells after drug treatment. Our homology modeling suggests that ligands bind PXR.1 more favorably, probably because of the presence of a key disordered loop region, which is missing in PXR.2. Yeast two-hybrid assays revealed that, even in the presence of ligand, the corepressors remain tightly bound to PXR.2, and coactivators are unable to bind at helix 12. In summary, PXR.2 can bind to PXREs but fails to transactivate target genes because ligands do not bind the ligand binding domain of PXR.2 productively, corepressors remain tightly bound, and coactivators are not recruited to PXR.2. The American Society for Pharmacology and Experimental Therapeutics ER -