New insights into the regulation of CYP2C9 gene expression: the role of the transcription factor GATA-4
| Autor: | Ellie Landman, Souren Mkrtchian, Yvonne Hofmann, Isa Cavaco, Jana Nekvindová, Magnus Ingelman-Sundberg, Rasmus Steen Pedersen, Jessica Mwinyi |
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| Přispěvatelé: | University of Zurich, Mwinyi, J |
| Rok vydání: | 2009 |
| Předmět: |
Chromatin Immunoprecipitation
3003 Pharmaceutical Science Activation Pharmaceutical Science 610 Medicine & health Electrophoretic Mobility Shift Assay Biology Transfection Gene Expression Regulation Enzymologic Cytochrome P4502C9 Mice Genes Reporter Cell Line Tumor GATA6 Transcription Factor Gene expression Transcriptional regulation Animals Humans Cyp19 Expression Electrophoretic mobility shift assay Binding site Promoter Regions Genetic Transcription factor Cytochrome P-450 CYP2C9 Pharmacology Binding Sites GATA2 Heart Hep G2 Cells Molecular biology Rat Granulosa-Cells GATA4 Transcription Factor DNA-Binding Proteins GATA2 Transcription Factor Nuclear Factor-4-Alpha Transporters 3004 Pharmacology Hepatocyte-Nuclear-Factor-4-Alpha Metabolism 10199 Clinic for Clinical Pharmacology and Toxicology embryonic structures Mutation Hepatocytes GATA transcription factor Aryl Hydrocarbon Hydroxylases Polymorphisms Chromatin immunoprecipitation Transcription Factors |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| ISSN: | 1521-009X |
| Popis: | CYP2C9 is an important drug-metabolizing enzyme that metabolizes, e. g., warfarin, antidiabetics, and antiphlogistics. However, the endogenous regulation of this enzyme is largely unknown. In this study, we examined the role of GATA transcription factors in the gene expression of CYP2C9. We investigated four putative GATA binding sites within the first 200 base pairs of CYP2C9 promoter at the positions I: -173/-170, II: -167/-164, III: -118/ -115, and IV: -106/-103. Luciferase activity driven by a wildtype CYP2C9 promoter construct was strongly up-regulated in Huh-7 cells upon cotransfection with expression plasmids for GATA-2 and GATA-4, whereas mutations introduced into GATA binding site III or I and II reduced this induction to a significant extent. Electrophoretic mobility shift assays revealed specific binding of GATA-4 and GATA-6 to the oligonucleotides containing GATA binding sites I and II. Furthermore, the association of GATA-4 with CYP2C9 promoter was confirmed by chromatin immunoprecipitation assays in HepG2 cells. Taken together, these data strongly suggest an involvement of liver-specific transcription factor GATA-4 in the transcriptional regulation of CYP2C9. Swedish Research Council; Stockholm County Council; Danish Agency of Science, Technology and Innovation; Lundbeck Foundation; Portuguese Foundation for Science and Technology [SFRH/BPD/34152/2006 IBB/CBME] |
| Databáze: | OpenAIRE |
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