Active ERK1/2 Protein Interacts with the Phosphorylated Nuclear Constitutive Active/androstane Receptor (CAR; NR1I3), Repressing Dephosphorylation and Sequestering CAR in the Cytoplasm

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2011 Aug 30

PMID 21873423

Citations 28

Authors

Makoto Osabe

Masahiko Negishi

Affiliations

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Abstract

The nuclear constitutive active/androstane receptor (CAR) is inactivated and sequestered in the cytoplasm when Thr-38 is phosphorylated. Here, we have demonstrated that activated ERK1/2 interacts with phosphorylated CAR to repress dephosphorylation of Thr-38. The phosphorylation-dependent interaction between CAR and ERK1/2 was examined by co-immunoprecipitation experiments of ectopically expressed FLAG-tagged CAR T38A and CAR T38D mutants with endogenous phospho-ERK1/2 in Huh-7 cells. Phospho-ERK1/2 coprecipitated only the phosphorylation-mimicking CAR T38D mutant; this coprecipitation was mediated by the interaction with the xenochemical response signal peptide near the C terminus of CAR. This interaction increased after EGF treatment and decreased after treatment with the MEK inhibitor U0126 as well as after knockdown of MEK1/2 by shRNA in Huh-7 cells. The phosphorylation levels of Thr-38 of CAR decreased in U0126-treated Huh-7 cells. Thus, activated ERK1/2 interacts with CAR and represses dephosphorylation of Thr-38, providing a cell signal-regulated mechanism for CAR activation.

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References

Dong B, Saha P, Huang W, Chen W, Abu-Elheiga L, Wakil S . Activation of nuclear receptor CAR ameliorates diabetes and fatty liver disease. Proc Natl Acad Sci U S A. 2009; 106(44):18831-6. PMC: 2773998. DOI: 10.1073/pnas.0909731106. View

Honkakoski P, Zelko I, Sueyoshi T, Negishi M . The nuclear orphan receptor CAR-retinoid X receptor heterodimer activates the phenobarbital-responsive enhancer module of the CYP2B gene. Mol Cell Biol. 1998; 18(10):5652-8. PMC: 109151. DOI: 10.1128/MCB.18.10.5652. View

Kamino H, Negishi M . The nuclear receptor constitutive active/androstane receptor arrests DNA-damaged human hepatocellular carcinoma Huh7 cells at the G2/M phase. Mol Carcinog. 2011; 51(2):206-12. PMC: 3242902. DOI: 10.1002/mc.20783. View

Koike C, Moore R, Negishi M . Extracellular signal-regulated kinase is an endogenous signal retaining the nuclear constitutive active/androstane receptor (CAR) in the cytoplasm of mouse primary hepatocytes. Mol Pharmacol. 2007; 71(5):1217-21. PMC: 2100393. DOI: 10.1124/mol.107.034538. View

Yamamoto Y, Moore R, Goldsworthy T, Negishi M, Maronpot R . The orphan nuclear receptor constitutive active/androstane receptor is essential for liver tumor promotion by phenobarbital in mice. Cancer Res. 2004; 64(20):7197-200. DOI: 10.1158/0008-5472.CAN-04-1459. View

Yamamoto Y, Moore R, Flavell R, Lu B, Negishi M . Nuclear receptor CAR represses TNFalpha-induced cell death by interacting with the anti-apoptotic GADD45B. PLoS One. 2010; 5(4):e10121. PMC: 2853562. DOI: 10.1371/journal.pone.0010121. View

Ueda A, Kakizaki S, Negishi M, Sueyoshi T . Residue threonine 350 confers steroid hormone responsiveness to the mouse nuclear orphan receptor CAR. Mol Pharmacol. 2002; 61(6):1284-8. DOI: 10.1124/mol.61.6.1284. View

Bauer D, Wolfram N, Kahl G, Hirsch-Ernst K . Transcriptional regulation of CYP2B1 induction in primary rat hepatocyte cultures: repression by epidermal growth factor is mediated via a distal enhancer region. Mol Pharmacol. 2004; 65(1):172-80. DOI: 10.1124/mol.65.1.172. View

Sueyoshi T, Kawamoto T, Zelko I, Honkakoski P, Negishi M . The repressed nuclear receptor CAR responds to phenobarbital in activating the human CYP2B6 gene. J Biol Chem. 1999; 274(10):6043-6. DOI: 10.1074/jbc.274.10.6043. View

10.

Wada T, Gao J, Xie W . PXR and CAR in energy metabolism. Trends Endocrinol Metab. 2009; 20(6):273-9. DOI: 10.1016/j.tem.2009.03.003. View

11.

Konno Y, Negishi M, Kodama S . The roles of nuclear receptors CAR and PXR in hepatic energy metabolism. Drug Metab Pharmacokinet. 2008; 23(1):8-13. DOI: 10.2133/dmpk.23.8. View

12.

Zelko I, Sueyoshi T, Kawamoto T, MOORE R, Negishi M . The peptide near the C terminus regulates receptor CAR nuclear translocation induced by xenochemicals in mouse liver. Mol Cell Biol. 2001; 21(8):2838-46. PMC: 86913. DOI: 10.1128/MCB.21.8.2838-2846.2001. View

13.

Kawamoto T, Sueyoshi T, Zelko I, MOORE R, Washburn K, Negishi M . Phenobarbital-responsive nuclear translocation of the receptor CAR in induction of the CYP2B gene. Mol Cell Biol. 1999; 19(9):6318-22. PMC: 84602. DOI: 10.1128/MCB.19.9.6318. View

14.

Kodama S, Koike C, Negishi M, Yamamoto Y . Nuclear receptors CAR and PXR cross talk with FOXO1 to regulate genes that encode drug-metabolizing and gluconeogenic enzymes. Mol Cell Biol. 2004; 24(18):7931-40. PMC: 515037. DOI: 10.1128/MCB.24.18.7931-7940.2004. View

15.

Rencurel F, Foretz M, Kaufmann M, Stroka D, Looser R, Leclerc I . Stimulation of AMP-activated protein kinase is essential for the induction of drug metabolizing enzymes by phenobarbital in human and mouse liver. Mol Pharmacol. 2006; 70(6):1925-34. DOI: 10.1124/mol.106.029421. View

16.

Mutoh S, Osabe M, Inoue K, Moore R, Pedersen L, Perera L . Dephosphorylation of threonine 38 is required for nuclear translocation and activation of human xenobiotic receptor CAR (NR1I3). J Biol Chem. 2009; 284(50):34785-92. PMC: 2787341. DOI: 10.1074/jbc.M109.048108. View

17.

Sueyoshi T, Negishi M . Phenobarbital response elements of cytochrome P450 genes and nuclear receptors. Annu Rev Pharmacol Toxicol. 2001; 41:123-43. DOI: 10.1146/annurev.pharmtox.41.1.123. View

18.

Shindo S, Numazawa S, Yoshida T . A physiological role of AMP-activated protein kinase in phenobarbital-mediated constitutive androstane receptor activation and CYP2B induction. Biochem J. 2006; 401(3):735-41. PMC: 1770843. DOI: 10.1042/BJ20061238. View

19.

Osabe M, Sugatani J, Takemura A, Yamazaki Y, Ikari A, Kitamura N . Expression of CAR in SW480 and HepG2 cells during G1 is associated with cell proliferation. Biochem Biophys Res Commun. 2008; 369(4):1027-33. DOI: 10.1016/j.bbrc.2008.02.140. View