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

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

doi:10.1016/j.bbrc.2003.12.027

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 journal › Article › peer-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 language	English
Pages (from-to)	922-925
Number of pages	4
Journal	Biochemical and Biophysical Research Communications
Volume	313
Issue number	4
DOIs	https://doi.org/10.1016/j.bbrc.2003.12.027
Publication status	Published - Jan 23 2004
Externally published	Yes

All Science Journal Classification (ASJC) codes

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2003.12.027

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@article{30e4f2c9d1244001b5424868619e4a99,

title = "A yeast one-hybrid system to detect methylation-dependent DNA-protein interactions",

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.",

keywords = "DNA methylation, Epigenetics, Kaiso, MBD1, Methyl CpG binding protein, Protein-DNA interaction, Yeast one-hybrid system",

author = "Feng, \{Shu Ying\} and Kazuhisa Ota and Yoichi Yamada and Norio Sawabu and Takashi Ito",

note = "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.",

year = "2004",

month = jan,

day = "23",

doi = "10.1016/j.bbrc.2003.12.027",

language = "English",

volume = "313",

pages = "922--925",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Elsevier B.V.",

number = "4",

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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.

KW - DNA methylation

KW - Epigenetics

KW - Kaiso

KW - MBD1

KW - Methyl CpG binding protein

KW - Protein-DNA interaction

KW - Yeast one-hybrid system

UR - https://www.scopus.com/pages/publications/0346098161

UR - https://www.scopus.com/pages/publications/0346098161#tab=citedBy

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 -

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

Abstract

All Science Journal Classification (ASJC) codes

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