Browse > Article
http://dx.doi.org/10.5352/JLS.2003.13.3.324

Effects of Constitutive Androstane Receptor (CAR) on PBRU Transactivation of CYP2B Gene in Different Culture Cell Types: Comparison Between Hep G2 and COS-cells  

민계식 (진주산업대학교 미생물공학과)
Publication Information
Journal of Life Science / v.13, no.3, 2003 , pp. 324-332 More about this Journal
Abstract
The objective of this study was to examine if transient transfection of CAR can transactivate CYP2B1 PBRU reporter gene in COS cells in which the endogenous CYP2B1 gene is not induced by PB. In non-transfeced cells of both Hep G2 and COS, the endogeneous expression of CAR was not detected by antibody against CAR. When cultured cells were transfected with CAR expression plasmid, mCAR1-GFP, both cell types expressed high levels of CAR protein and could allow to examine the effect of CAR in PBRU transactivation. Both cell types expressed endogenous RXR and transfection of RXR expression plasmid dramatically increased its protein expression. Whereas CAR transactivated PBRU2C1Luciferase about 12 fold as compared to 2C1Luciferase in Hep G2 cells, it did not stimulate the luciferase activity of the PBRU reporter gene in COS cells. These results indicate that Hep G2 cells can respond to CAR differently from COS cells, and suggest that factors other than CAR and RXR may be required in inducing PBRU activation and the expression of these factors may be different between liver and kidney.
Keywords
Constitutive Androstane Receptor; PBRU; Transactivation; Hep G2; COS;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Wei, P., J. Zhang, M. Egan-Hafley, S. Liang and D. D. Moore. 2000. The nuclear receptor CAR mediates specific xenobiotic induction of drug metabolism. Nature. 407, 920-923   DOI   ScienceOn
2 Sueyoshi, T., T. Kawamoto, I. Zelko, P. Honkakoski and M. Negishi. 1999. The repressed nuclear receptor CAR responds to phenobarbital in activating the human CYP2B6 gene. J. Biol. Chem. 274, 6043-6046   DOI   ScienceOn
3 Sueyoshi, T. and M. Negishi. 2001. Phenobarbital response elements of cytochrome P450 genes and nuclear receptors. Annu. Rev. Pharmacol. Toxicol. 41, 123-143   DOI   ScienceOn
4 Suwa, Y, Y Mizukami, K. Sogawa and Y. FujiiKuriyama. 1985. Gene structure of a major form of phenobarbital-inducible cytochrome P-450 in rat liver. J. Biol. Chem. 260, 7980-7984
5 Trottier, E., A. Belzil, C. Stoltz and A. Anderson. 1995. Localization of a phenobarbital-responsive el-ement (PBRE) in the 5'-flanking region of the rat CYP2B2 gene. Gene 158, 263-268   DOI   ScienceOn
6 Tzameli, I., P. Pissios, E. G. Schuetz and D. D. Moore. 2000. The xenobiotic compound 1,4-bis[2-(3,5dichloropyridyloxy)] benzene is an agonist ligand for the nuclear receptor CAR. Mol. Cell Bioi. 20, 2951-2958   DOI   ScienceOn
7 Waxman, D. J. 1999. P450 gene induction by struc-turally diverse xenochemicals: central role of nuclear receptors CAR, PXR, and PPAR. Archives of Biochemistry and Biophysics 369, 11-23   DOI   ScienceOn
8 Waxman, D. J. and L. Azaroff. 1992. Phenobarbital induction of cytochrome P-450 gene expression. Biochem. J. 281, 577-592
9 Negishi, M. and T. Sueyoshi. 2001. Phenobarbital response elements of Cytochrome P450 genes and nuclear receptors. Annu. Rev. Pharmacol. Toxicol. 41,123-143   DOI   ScienceOn
10 Park, Y., H. Li and B. Kemper. 1996. Phenobarbital induction mediated by a distal CYP2B2 sequence in rat liver transiently transfected in situ. J. Biol. Chem. 271, 23725-23728   DOI   ScienceOn
11 Ramsden, R., N. B. Beck, K. M. Sommer and C. J. Omiecinski. 1999. Phenobarbital responsiveness con-ferred by the 5-flanking region of the rat CYP2B2 gene in transgenic mice. Gene. 228, 169-179   DOI   ScienceOn
12 Ramsden, R., K. M. Sommer and C. J. Omiecinski. 1993. Phenobarbital induction and tissue-specific ex-pression of the rat CYP2B2 gene in transgenic mice. J. BioI. Chem. 268, 21722-21726
13 Savas, U., K. J. Griffin and E. F. Johnson. 1999. Molecular mechanisms of cytochrome P-450 induction by xenobiotics: An expanded role for nuclear hormone receptors. Molecular Pharmacology. 56, 851-857
14 Shervington, A. 1999. CYP2B2 gene expression and phenobarbital induction in kidneys using an in vitro transcription system. Biochem. Mol. BioI. Int. 47, 233-237
15 Min, G., J. K. Kemper and B. Kemper. 2002. Glucocorticoid receptor-interacting protein 1 mediates ligand-independent nuclear translocation and activation of constitutive androstane receptor in uiuo. J. BioI. Chem. 277, 26356-26363   DOI   ScienceOn
16 Strobel, H. W., A. Y H. Lu, J. Heidma and M. J. Coon. 1970. Phosphotidylcholine requirement in the enzymatic reduction of hemoprotein P-450 and in fatty acid, hydrocarbon, and drug hydroxylation. J. Biol. Chem. 245, 4851-4954
17 Kim, J., I. Rivera-Rivera and B. Kemper. 2000. Tissue-specific chromatin structure of the CYP2B1 phenobar-bital-responsive unit and proximal promotor and modulation by phenobarbital. Nucleic Acids Res. 28, 1126-1132   DOI   ScienceOn
18 Liu, S., Y. Park, I. Rivera-Rivera, H. Li and B. Kemper. 1998. Nuclear factor-1 motif and redundant regulatory elements comprise phenobarbital-responsive enhancer in CYP2B1/2. DNA Cell Bioi. 17, 461-470   DOI   ScienceOn
19 Moore, D. and I. Tzameli. 2001. Role reversal: new insights from new ligands for the xenobiotic receptor CAR. TRENDS in Endocrinology and Metabolism 12, 7-10   DOI   ScienceOn
20 Moore, L. B., D. J. Parks, S. A. Jones, R. K. Bledsoe, T. G. Consler, J. B. Stimmel, B. Goodwin, C. Liddle, S. G. Blanchard, T. M. Willson, J. L. Collins and S. A. Kliewer. 2000. Orphan nuclear receptors con-stitutive androstane receptor and pregnene X receptor share xenobiotic and steroid ligands. J. Biol. Chern. 275, 15122-15127   DOI   ScienceOn
21 Mueller, G. C. and J. A. Miller. 1949. The reductive cleavage of 4-demethylaminoazobenzene by rat liver: The intracellular distribution of the enzyme system and the requirement for triphosphopyridine nucleotide. J. BioI. Chem. 180, 1125-1136
22 Negishi, M. and P. Honkakoski. 2000. Regulation of cytochrome P450 (CYP) genes by nuclear receptors. Biochem. J. 347, 321-337   DOI   ScienceOn
23 Kim, J. and B. Kemper. 1997. Phenobarbital alters protein binding to the CYP2Bl/2 phenobarbital-responsive unit in native chromatin. J. Bioi. Chem. 272, 29423-29426   DOI   ScienceOn
24 Jean, A., A. Reiss, M. Desrochers, S. Dubois, E. Trottier, Y. Trottier, L. Wirtanen, M. Adesnik, D. J. Waxman and A. Anderson. 1994. Rat liver cyto-chrome P450 2B3: structure of the CYP2B3 gene and immunological identification of a constitutive P450 2B3-like protein in rat liver. DNA Cell BioI. 13, 781-792   DOI   ScienceOn
25 Johnson, E. F., K. J. Griffin and U. S. Keith. 1999. Molecular mechanisms of cytochrome P-450 induction by xenobiotics: An expanded role for nuclear hormone receptors. Molecular Pharmacology. 56, 851-857
26 Kawamoto, T., T. Sueyoshi, I. Zelko, R. Moore, K. Washburn and M. Negish. 1999. Phenobarbital responsive nuclear translocation of the receptor CAR in induction of the CYP2B gene. Mol. Cell Bioi. 19, 6318-6322
27 Honkakoski, P., R. Moore, K. Washburn and M. Negishi. 1988. Activation by diverse xenochemicals of the 51-base pair phenobarbital-response enhancer module in the CYP2B10 gene. Mol. Pharmacal. 53, 597-601
28 Kim, J., G. Min and B. Kemper. 2001. Chromatin assembly enhances binding to the CYP2B1 phenobar-bital-responsive unit (PBRD) of nuclear factor-1, which binds simultaneously with constitutive andro-stane receptor (CAR)/retinoid $\times$receptor (RXR) and enhances CAR/RXR-mediated activation of the PBRU. J. Biol. Chem. 276, 7559-7567   DOI   ScienceOn
29 Forman, B. M., I. Tzameli, H. S. Choi, J. Chen, D. Simha, W. Seol, R. M. Evans and D. D. Moore. 1998. Androstane metabolites bind to and deactivate the nuclear receptor CAR-beta. Nature. 395, 612-615   DOI   ScienceOn
30 Honkakoski, P., A. Kojo and M. A. Lang. 1992. Reg-ulation of the mouse liver cytochrome P450 2B subfamily by sex hormones and phenobarbital. Biochemical. J. 285, 979-983
31 Honkakoski, P. and M. Negishi. 1997. Character-ization of a phenobarbital-responsive enhancer mod-ule in mouse P450 Cyp2blO gene. J. BioI. Chem. 272, 14943-14949   DOI   ScienceOn
32 Honkakoski, P. and M. Negishi. 2000. Regulation of cytochrome P450 (CYP) genes by nuclear receptors. Biochem. J. 347, 321-337   DOI   ScienceOn
33 Choi, H-S., M. Chung, I. Tzameli, D. Simha, Y-K. Lee, W. Seol and D. Moore. 1997. Differential trans-activation by two isoforms of the orphan nuclear hormone receptor CAR. J. BioI. Chem. 272, 23565-23571   DOI   ScienceOn
34 Honkakoski, P., I. Zelko, T. Sueyoshi and M. Negishi. 1998. The nuclear orphan receptor CAR-retinoid X receptor heterodimer activates the phenobarbital-responsive enhancer module of the CYP2B gene. Mol. Cell Bioi. 18, 5652-5658
35 Bhagwat, S. V., B. C. Leelavathi, S. K. Shankar, M. R. Boyd and V. avindranath. 1995. Cytochrome P450 and associated monooxygenase activities in the rat and human spinal cord-induction, immunological char-acterization, and immunocytochemical localization. Neurosci. 68, 593-601   DOI   ScienceOn
36 Brodie, B. B., J. Axelrod, J. R. Cooper, L. Gaudette, B. N. LaDu, C. Mitoma and S. Udenfriend. 1955. Detox-ication of drugs and other foreign compounds by liver microsomes. Science 121, 603-604   DOI
37 Conney, A. H., C. Davison, R. Gasten and J. J. Burns. 1960. Adaptive increase in drug-metabolizing enzymes induced by phenobarbital and other drugs. J. Pharmacal. Exp. Therap. 130, 1-8
38 Baes, M., T. Gulick, H. S. Choi, M. G. Martinoli, D. Simha and D. D. Moore. 1994. A new orphan member of the nuclear hormone receptor superfamily that interacts with a subset of retinoic acid response ele-ments. Mol. Cell Bioi. 14, 1544-1551
39 Cooper, D. Y., S. Levine, S. Narasimhulu, O. Ro-senthal and R. W. Estabrook. 1965. Photochemical action spectrum of the terminal oxidase of 'mixed function oxidase system. Science. 147, 400-402   DOI   ScienceOn
40 Axelrod, J. 1955. The enzymatic deamination of am-phetamine (benzedrine). J. Bioi. Chem. 214, 753-763