Regulation of human CYP2C9 expression by electrophilic stress involves activator protein 1 activation and DNA looping

Ngome L. Makia, Sailesh Surapureddi, K. Monostory, Russell A. Prough, Joyce A. Goldstein

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Cytochrome P450 (CYP)2C9 and CYP2C19 are important human enzymes that metabolize therapeutic drugs, environmental chemicals, and physiologically important endogenous compounds. Initial studies using primary human hepatocytes showed induction of both the CYP2C9 and CYP2C19 genes by tertbutylhydroquinone (tBHQ). As a pro-oxidant, tBHQ regulates the expression of cytoprotective genes by activation of redox-sensing transcription factors, such as the nuclear factor E2-related factor 2 (Nrf2) and members of the activator protein 1 (AP-1) family of proteins. The promoter region of CYP2C9 contains two putative AP-1 sites (TGAGTCA) at positions -2201 and -1930, which are also highly conserved in CYP2C19. The CYP2C9 promoter is activated by ectopic expression of cFos and JunD, whereas Nrf2 had no effect. Using specific kinase inhibitors formitogen-activated protein kinase, we showed that extracellular signal-regulated kinase and Jun N-terminal kinase are essential for tBHQ-induced expression of CYP2C9. Electrophoretic mobility shift assays demonstrate that cFos distinctly interacts with the distal AP-1 site and JunD with the proximal site. Because cFos regulates target genes as heterodimers with Jun proteins, we hypothesized that DNA looping might be required to bring the distal and proximal AP-1 sites together to activate the CYP2C9 promoter. Chromosome conformation capture analyses confirmed the formation of a DNA loop in the CYP2C9 promoter, possibly allowing interaction between cFos at the distal site and JunD at the proximal site to activate CYP2C9 transcription in response to electrophiles. These results indicate that oxidative stress generated by exposure to electrophilic xenobiotics and metabolites induces the expression of CYP2C9 and CYP2C19 in human hepatocytes. U.S. Government work not protected by U.S. copyright.

Original languageEnglish
Pages (from-to)125-137
Number of pages13
JournalMolecular Pharmacology
Volume86
Issue number2
DOIs
Publication statusPublished - 2014

Fingerprint

Transcription Factor AP-1
DNA
NF-E2-Related Factor 2
Hepatocytes
Phosphotransferases
Cytochrome P-450 CYP2C9
Extracellular Signal-Regulated MAP Kinases
Electrophoretic Mobility Shift Assay
Xenobiotics
Genetic Promoter Regions
Cytochrome P-450 Enzyme System
Protein Kinases
Transcriptional Activation
Genes
Oxidation-Reduction
Reactive Oxygen Species
Proteins
Oxidative Stress
Transcription Factors
Chromosomes

ASJC Scopus subject areas

  • Pharmacology
  • Molecular Medicine

Cite this

Regulation of human CYP2C9 expression by electrophilic stress involves activator protein 1 activation and DNA looping. / Makia, Ngome L.; Surapureddi, Sailesh; Monostory, K.; Prough, Russell A.; Goldstein, Joyce A.

In: Molecular Pharmacology, Vol. 86, No. 2, 2014, p. 125-137.

Research output: Contribution to journalArticle

Makia, Ngome L. ; Surapureddi, Sailesh ; Monostory, K. ; Prough, Russell A. ; Goldstein, Joyce A. / Regulation of human CYP2C9 expression by electrophilic stress involves activator protein 1 activation and DNA looping. In: Molecular Pharmacology. 2014 ; Vol. 86, No. 2. pp. 125-137.
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