Constitutive androstane receptor 1 is constitutively bound to chromatin and ‘primed’ for transactivation in hepatocytes

Michael McMahon, Shaohong Ding, Lourdes Acosta-Jimenez, Remi Terranova, Marie-Apolline Gerard, Antonio Vitobello, Jonathan G. Moggs, Colin J. Henderson, C. Ronald Wolf (Lead / Corresponding author)

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Abstract

The constitutive androstane receptor (CAR) is a xenobiotic sensor expressed in hepatocytes that activates genes involved in drug metabolism, lipid homeostasis, and cell proliferation. Much progress has been made in understanding the mechanism of activation of human CAR by drugs and xenobiotics. However, many aspects of the activation pathway remain to be elucidated. In this report, we have used viral constructs to express human CAR, its splice variants, and mutant CAR forms in hepatocytes from Car-/- mice in vitro and in vivo. We demonstrate CAR expression rescued the ability of Car-/- hepatocytes to respond to a wide range of CAR activators including phenobarbital. Additionally, two major splice isoforms of human CAR, CAR2 and CAR3, were inactive with almost all the agents tested. In contrast to the current model of CAR activation, ectopic CAR1 is constitutively localised in the nucleus and is loaded onto Cyp2b10 gene in the absence of an inducing agent. In studies to elucidate the role of threonine T38 in CAR regulation, we found that the T38D mutant was inactive even in the presence of CAR activators. However, the T38A mutant was activated by CAR inducers, showing that T38 is not essential for CAR activation. Also, using the inhibitor erlotinib, we could not confirm a role for the epidermal growth factor receptor in CAR regulation. Our data suggest that CAR is constitutively bound to gene regulatory regions and is regulated by exogenous agents through a mechanism which involves protein phosphorylation in the nucleus.
Original languageEnglish
Pages (from-to)97-105
Number of pages9
JournalMolecular Pharmacology
Volume95
Issue number1
Early online date25 Oct 2018
DOIs
Publication statusPublished - 1 Jan 2019

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Transcriptional Activation
Chromatin
Hepatocytes
Xenobiotics
constitutive androstane receptor
Genes
Nucleic Acid Regulatory Sequences
Threonine
Phenobarbital
Lipid Metabolism
Epidermal Growth Factor Receptor
Pharmaceutical Preparations
Protein Isoforms
Homeostasis
Phosphorylation
Cell Proliferation

Keywords

  • Drug metabolism
  • Liver
  • Nuclear receptors
  • Protein phosphorylation
  • Regulation of gene expression

Cite this

McMahon, Michael ; Ding, Shaohong ; Acosta-Jimenez, Lourdes ; Terranova, Remi ; Gerard, Marie-Apolline ; Vitobello, Antonio ; Moggs, Jonathan G. ; Henderson, Colin J. ; Wolf, C. Ronald. / Constitutive androstane receptor 1 is constitutively bound to chromatin and ‘primed’ for transactivation in hepatocytes. In: Molecular Pharmacology. 2019 ; Vol. 95, No. 1. pp. 97-105.
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abstract = "The constitutive androstane receptor (CAR) is a xenobiotic sensor expressed in hepatocytes that activates genes involved in drug metabolism, lipid homeostasis, and cell proliferation. Much progress has been made in understanding the mechanism of activation of human CAR by drugs and xenobiotics. However, many aspects of the activation pathway remain to be elucidated. In this report, we have used viral constructs to express human CAR, its splice variants, and mutant CAR forms in hepatocytes from Car-/- mice in vitro and in vivo. We demonstrate CAR expression rescued the ability of Car-/- hepatocytes to respond to a wide range of CAR activators including phenobarbital. Additionally, two major splice isoforms of human CAR, CAR2 and CAR3, were inactive with almost all the agents tested. In contrast to the current model of CAR activation, ectopic CAR1 is constitutively localised in the nucleus and is loaded onto Cyp2b10 gene in the absence of an inducing agent. In studies to elucidate the role of threonine T38 in CAR regulation, we found that the T38D mutant was inactive even in the presence of CAR activators. However, the T38A mutant was activated by CAR inducers, showing that T38 is not essential for CAR activation. Also, using the inhibitor erlotinib, we could not confirm a role for the epidermal growth factor receptor in CAR regulation. Our data suggest that CAR is constitutively bound to gene regulatory regions and is regulated by exogenous agents through a mechanism which involves protein phosphorylation in the nucleus.",
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Constitutive androstane receptor 1 is constitutively bound to chromatin and ‘primed’ for transactivation in hepatocytes. / McMahon, Michael; Ding, Shaohong; Acosta-Jimenez, Lourdes; Terranova, Remi; Gerard, Marie-Apolline; Vitobello, Antonio; Moggs, Jonathan G.; Henderson, Colin J.; Wolf, C. Ronald (Lead / Corresponding author).

In: Molecular Pharmacology, Vol. 95, No. 1, 01.01.2019, p. 97-105.

Research output: Contribution to journalArticle

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AU - Ding, Shaohong

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AU - Gerard, Marie-Apolline

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AU - Wolf, C. Ronald

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