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PMID: 14555760 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Methylated DNA-binding domain 1 and methylpurine-DNA glycosylase link transcriptional repression and DNA repair in chromatin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 100 ·No. 22 ·2003-10-28 ·Pages 12859-64

Watanabe S, Ichimura T, Fujita N, Tsuruzoe S, Ohki I, Shirakawa M, Kawasuji M, Nakao M

Abstract

The methyl-CpG dinucleotide containing a symmetrical 5-methylcytosine (mC) is involved in gene regulation and genome stability. We report here that methylation-mediated transcriptional repressor methylated DNA-binding domain 1 (MBD1) interacts with methylpurine-DNA glycosylase (MPG), which excises damaged bases from substrate DNA. MPG itself actively represses transcription and has a synergistic effect on gene silencing together with MBD1. Chromatin immunoprecipitation analysis reveals the molecular movement of MBD1 and MPG in vivo: (i) The MBD1-MPG complex normally exists on the methylated gene promoter; (ii) treatment of cells with alkylating agent methylmethanesulfonate (MMS) induces the dissociation of MBD1 from the methylated promoter, and MPG is located on both methylated and unmethylated promoters; and (iii) after completion of the repair, the MBD1-MPG complex is restored on the methylated promoter. Mobility-shift and structural analyses show that the MBD of MBD1 binds a methyl-CpG pair (mCpG x mCpG) but not the methyl-CpG pair containing a single 7-methylguanine (N) (mCpG x mCpN) that is known as one of the major lesions caused by MMS. We further demonstrate that knockdown of MBD1 by specific small interfering RNAs significantly increases cell sensitivity to MMS. These data suggest that MBD1 cooperates with MPG for transcriptional repression and DNA repair. We hypothesize that MBD1 functions as a reservoir for MPG and senses the base damage in chromatin

MeSH Terms
Base Sequence Binding Sites Chromatin/genetics Cross-Linking Reagents DNA Glycosylases/chemistry,metabolism DNA Methylation DNA Repair/genetics DNA, Neoplasm/chemistry,metabolism Dinucleoside Phosphates/chemistry,metabolism Gene Expression Regulation, Neoplastic Glutathione Transferase/genetics HeLa Cells Humans Plasmids RNA, Small Interfering/genetics Recombinant Fusion Proteins/metabolism Transcription, Genetic Transfection Tumor Cells, Cultured
Chemicals
Chromatin Cross-Linking Reagents DNA, Neoplasm Dinucleoside Phosphates RNA, Small Interfering Recombinant Fusion Proteins cytidylyl-3'-5'-guanosine Glutathione Transferase DNA Glycosylases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Watanabe Sugiko
Department of Regeneration Medicine, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan.
Ichimura Takaya
Fujita Naoyuki
Tsuruzoe Shu
Ohki Izuru
Shirakawa Masahiro
Kawasuji Michio
Nakao Mitsuyoshi
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.2131819100
Published
2003-10-28
Epub
2003-00-10
Pages
12859-64
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC240709
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.2131819100 PMC Full Text View journal details
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