Sequence diversity and functional characterization of the 5′-regulatory region of human CYP2C19

Million Arefayene, Todd Skaar, Xiaojiong Zhao, James M. Rae, Jose E. Tanus-Santos, Ulrich Brinkmann, Ilka Brehm, Ulrike Salat, Anne Nguyen, Zeruesenay Desta, David A. Flockhart

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Cytochrome P4502C19 (CYP2C19) plays an important role in drug biotransformation and has been shown to be genetically polymorphic. While polymorphisms in the coding region that have large effects on activity are well described, until recently, a lack of knowledge of the promoter sequence has hindered efforts to study it. Genetic variants in the promoter region have not been described and factors that influence its gene expression via promotor regulation remain largely undefined. We have cloned and sequenced 1.8 kb of the human CYP2C19 promoter. This promoter contains a number of putative transcription factor sites, including HepG2-specific factor 1, glucocorticoid response element, estrogen receptor element, constitutive androstane receptor and peroxisome proliferator-activated receptor. Sequencing of DNA obtained from 67 individuals identified eight single nucleotide polymorphisms within this region. No sequence of a known human pregnane X receptor response element was found in this section of the CYP2C19 promoter, despite the known effect of rifampin on the expression of this gene. A plasmid containing the 1.8-kb CYP2C19 promoter coupled to a luciferase reporter gene has been constructed and demonstrated to be functional and sensitive to induction by omeprazole in HuH7 cells. Nested deletions of CYP2C19 promoter were generated and the ability of serial promoter deletion constructs to activate luciferase expression in the HepG2 cell line was analysed. These data make possible future studies to elucidate the molecular mechanisms by which CYP2C19 can be induced in clinical settings and the consequences of genetic variability in its promoter.

Original languageEnglish
Pages (from-to)199-206
Number of pages8
JournalPharmacogenetics
Volume13
Issue number4
DOIs
StatePublished - Apr 2003

Fingerprint

Nucleic Acid Regulatory Sequences
Cytochromes
Response Elements
Luciferases
Gene Expression
Peroxisome Proliferator-Activated Receptors
Omeprazole
Hep G2 Cells
Rifampin
Biotransformation
DNA Sequence Analysis
Reporter Genes
Genetic Promoter Regions
Estrogen Receptors
Glucocorticoids
Single Nucleotide Polymorphism
human CYP2C19 protein
Plasmids
Transcription Factors
Cell Line

Keywords

  • CYP2C19
  • Omeprazole
  • Polymorphism
  • Promoter
  • Rifampin
  • Sequence
  • Transcription factor

ASJC Scopus subject areas

  • Genetics
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Arefayene, M., Skaar, T., Zhao, X., Rae, J. M., Tanus-Santos, J. E., Brinkmann, U., ... Flockhart, D. A. (2003). Sequence diversity and functional characterization of the 5′-regulatory region of human CYP2C19. Pharmacogenetics, 13(4), 199-206. https://doi.org/10.1097/00008571-200304000-00004

Sequence diversity and functional characterization of the 5′-regulatory region of human CYP2C19. / Arefayene, Million; Skaar, Todd; Zhao, Xiaojiong; Rae, James M.; Tanus-Santos, Jose E.; Brinkmann, Ulrich; Brehm, Ilka; Salat, Ulrike; Nguyen, Anne; Desta, Zeruesenay; Flockhart, David A.

In: Pharmacogenetics, Vol. 13, No. 4, 04.2003, p. 199-206.

Research output: Contribution to journalArticle

Arefayene, M, Skaar, T, Zhao, X, Rae, JM, Tanus-Santos, JE, Brinkmann, U, Brehm, I, Salat, U, Nguyen, A, Desta, Z & Flockhart, DA 2003, 'Sequence diversity and functional characterization of the 5′-regulatory region of human CYP2C19', Pharmacogenetics, vol. 13, no. 4, pp. 199-206. https://doi.org/10.1097/00008571-200304000-00004
Arefayene, Million ; Skaar, Todd ; Zhao, Xiaojiong ; Rae, James M. ; Tanus-Santos, Jose E. ; Brinkmann, Ulrich ; Brehm, Ilka ; Salat, Ulrike ; Nguyen, Anne ; Desta, Zeruesenay ; Flockhart, David A. / Sequence diversity and functional characterization of the 5′-regulatory region of human CYP2C19. In: Pharmacogenetics. 2003 ; Vol. 13, No. 4. pp. 199-206.
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