Regulation of the promoter activity of interferon regulatory factor-7 gene. Activation by interferon snd silencing by hypermethylation

J Biol Chem. 2000 Oct 13;275(41):31805-12. doi: 10.1074/jbc.M005288200.

Abstract

The molecular mechanism by which virus induces expression of the early inflammatory genes has not yet been completely elucidated. Previous studies indicated that the virus-mediated transcription of type I interferon (IFN) genes required activation of two members of IFN regulatory factor (IRF) family, IRF-3 and IRF-7, where the expression of IRF-7 was found to be indispensable for the induction of IFNA genes. To determine the factors that regulate expression of IRF-7 gene, as well as its inducibility by type I IFNs, we have isolated and characterized the promoter and first intron of the human IRF-7 gene. This region shows a presence of two potential interferon-sensitive response elements (ISRE/IRF-E). However, only the ISRE present in the first intron was functional and conferred interferon inducibility in a transient transfection assay. Using a pull-down assay with an oligodeoxynucleotide corresponding to this ISRE immobilized to magnetic beads, we have demonstrated that this ISRE binds ISGF3 complex and IRF-1 from the extract of IFN-treated cells but not from the untreated cells. We have further shown that the previously observed lack of expression of IRF-7 in 2fTGH fibrosarcoma cell line, correlated with hypermethylation of the CpG island in the human IRF-7 promoter. The repression of the promoter activity was relieved by treatment with DNA methyltransferase inhibitor 5-aza-deoxycytidine. In vitro methylation of IRF-7 promoter silenced IRF-7 directed expression of luciferase gene in HeLa cells that express endogenous IRF-7 gene. Whether silencing of IRF-7 by methylation is instrumental for the process of tumorigenesis remains to be determined.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Azacitidine / analogs & derivatives
  • Azacitidine / pharmacology
  • Base Sequence
  • Cloning, Molecular
  • CpG Islands / genetics
  • DNA / genetics
  • DNA / metabolism
  • DNA Methylation / drug effects*
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Decitabine
  • Gene Silencing / drug effects*
  • Humans
  • Interferon Regulatory Factor-7
  • Interferon-Stimulated Gene Factor 3
  • Interferon-Stimulated Gene Factor 3, gamma Subunit
  • Interferon-alpha / pharmacology*
  • Introns / genetics
  • Molecular Sequence Data
  • Mutation / genetics
  • Oligodeoxyribonucleotides / genetics
  • Oligodeoxyribonucleotides / metabolism
  • Promoter Regions, Genetic / genetics*
  • Protein Binding
  • Response Elements / genetics
  • Transcription Factors / metabolism
  • Transcriptional Activation / drug effects*
  • Tumor Cells, Cultured

Substances

  • DNA-Binding Proteins
  • IRF7 protein, human
  • IRF9 protein, human
  • Interferon Regulatory Factor-7
  • Interferon-Stimulated Gene Factor 3
  • Interferon-Stimulated Gene Factor 3, gamma Subunit
  • Interferon-alpha
  • Oligodeoxyribonucleotides
  • Transcription Factors
  • Decitabine
  • DNA
  • Azacitidine

Associated data

  • GENBANK/AF277159