A yeast one-hybrid system to detect methylation-dependent DNA-protein interactions

Shu Ying Feng, Kazuhisa Ota, Yoichi Yamada, Norio Sawabu, Takashi Ito

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

We developed a method for site-selective CpG methylation of the budding yeast genome. The method recruits LexA-fused M.SssI DNA methyltransferase to LexA operator sequences integrated adjacent to the target site. Microarray analysis of methylated DNAs indicated that the tethered enzyme selectively methylates the region around the target site. Exploiting this method to methylate bait DNA in the one-hybrid system, we demonstrated methylation-dependent DNA binding of methyl-CpG binding proteins, MBD1 and Kaiso, in vivo. This methylation-dependent one-hybrid system would provide a versatile tool for the search and analysis of proteins that recognize methylated DNA to participate in epigenetic regulation.

Original languageEnglish
Pages (from-to)922-925
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume313
Issue number4
DOIs
Publication statusPublished - Jan 23 2004
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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