Phenobarbital-Responsive Nuclear Translocation of the Receptor CAR in Induction of the CYP2B Gene

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Molecular and Cellular Biology, September 1999, p. 6318-6322, Vol. 19, No. 9
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Phenobarbital-Responsive Nuclear Translocation of the Receptor CAR in Induction of the CYP2B Gene

Takeshi Kawamoto, Tatsuya Sueyoshi, Igor Zelko, Rick Moore, Kimberly Washburn, and Masahiko Negishi^*

Pharmacogenetics Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709

Received 16 April 1999/Returned for modification 1 June 1999/Accepted 16 June 1999

The constitutively active receptor (CAR) transactivates a distal enhancer called the phenobarbital (PB)-responsive enhancermodule (PBREM) found in PB-inducible CYP2B genes. CAR dramaticallyincreases its binding to PBREM in livers of PB-treated mice. Wehave investigated the cellular mechanism of PB-induced increaseof CAR binding. Western blot analyses of mouse livers revealedan extensive nuclear accumulation of CAR following PB treatment.Nuclear contents of CAR perfectly correlate with an increase ofCAR binding to PBREM. PB-elicited nuclear accumulation of CARappears to be a general step regulating the induction of CYP2Bgenes, since treatments with other PB-type inducers result inthe same nuclear accumulation of CAR. Both immunoprecipitationand immunohistochemistry studies show cytoplasmic localizationof CAR in the livers of nontreated mice, indicating that CAR translocatesinto nuclei following PB treatment. Nuclear translocation of CARalso occurs in mouse primary hepatocytes but not in hepatocytestreated with the protein phosphatase inhibitor okadaic acid. Thus,the CAR-mediated transactivation of PBREM in vivo becomes PB responsivethrough an okadaic acid-sensitive nuclear translocationprocess.

^* Corresponding author. Mailing address: Laboratory of Reproductive and Developmental Toxicology, National Institute of EnvironmentalHealth Sciences, National Institutes of Health, Research TrianglePark, NC 27709. Phone: (919) 541-2404. Fax: (919) 541-0696. E-mail:negishi{at}niehs.nih.gov.

Molecular and Cellular Biology, September 1999, p. 6318-6322, Vol. 19, No. 9
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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Li-Masters, T., Morgan, E. T. (2001). Effects of Bacterial Lipopolysaccharide on Phenobarbital-Induced CYP2B Expression in Mice. Drug Metab. Dispos. 29: 252-257 [Abstract] [Full Text]
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Moore, L. B., Parks, D. J., Jones, S. A., Bledsoe, R. K., Consler, T. G., Stimmel, J. B., Goodwin, B., Liddle, C., Blanchard, S. G., Willson, T. M., Collins, J. L., Kliewer, S. A. (2000). Orphan Nuclear Receptors Constitutive Androstane Receptor and Pregnane X Receptor Share Xenobiotic and Steroid Ligands. J. Biol. Chem. 275: 15122-15127 [Abstract] [Full Text]
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