TY - JOUR
T1 - A yeast one-hybrid system to detect methylation-dependent DNA-protein interactions
AU - Feng, Shu Ying
AU - Ota, Kazuhisa
AU - Yamada, Yoichi
AU - Sawabu, Norio
AU - Ito, Takashi
N1 - Funding Information:
We thank R.J. Roberts and S. Stickel (New England Biolabs) for their generous gift of M.SssI plasmid. This work is, in part, supported by grants from Ministry of Education, Culture, Sports, Science and Technology, Japan.
PY - 2004/1/23
Y1 - 2004/1/23
N2 - 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.
AB - 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.
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U2 - 10.1016/j.bbrc.2003.12.027
DO - 10.1016/j.bbrc.2003.12.027
M3 - Article
C2 - 14706630
AN - SCOPUS:0346098161
SN - 0006-291X
VL - 313
SP - 922
EP - 925
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 4
ER -