Targeted methylation of CMV and E1A viral promoters

Chia Chen Hsu, Hsin Pai Li, Yu Hung Hung, Yu Wei Leu, Wu Hsiung Wu, Feng Sheng Wang, Kuan Der Lee, Pey Jium Chang, Chi Sheng Wu, Yen Jung Lu, Tim H M Huang, Yu Sun Chang, Shu Huei Hsiao

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

DNA methylation is a gene-silencing and host defense system that can down-regulate viral gene expression in mammalian cells. An established targeted DNA methylation method was used to demonstrate that genome-integrated CMV and adenovirus type 5 E1A promoters were hypermethylated after MCF7 and HEK293 cells were transfected with in vitro methylated viral promoter fragments. In both cases, the targeted methylation-induced gene silencing could be reversed by addition of 5-aza-2'-deoxycytidine, confirming that the CMV and E1A promoters are regulated by DNA methylation. The kinetics of the targeted DNA methylation was determined using a reporter system in live cells. In conclusion, targeted DNA methylation is able to efficiently silence susceptible viral promoters and provides an alternative strategy to study the impact of loci-specific DNA methylation in viral gene expression.

Original languageEnglish
Pages (from-to)228-234
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume402
Issue number2
DOIs
Publication statusPublished - Nov 12 2010
Externally publishedYes

Keywords

  • DNA methylation
  • Epigenetics
  • Targeted DNA methylation
  • Two-component EGFP reporter system
  • Viral promoter

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

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  • Cite this

    Hsu, C. C., Li, H. P., Hung, Y. H., Leu, Y. W., Wu, W. H., Wang, F. S., Lee, K. D., Chang, P. J., Wu, C. S., Lu, Y. J., Huang, T. H. M., Chang, Y. S., & Hsiao, S. H. (2010). Targeted methylation of CMV and E1A viral promoters. Biochemical and Biophysical Research Communications, 402(2), 228-234. https://doi.org/10.1016/j.bbrc.2010.09.131