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PMID:17212651

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Citation

Ikegami, K, Iwatani, M, Suzuki, M, Tachibana, M, Shinkai, Y, Tanaka, S, Greally, JM, Yagi, S, Hattori, N and Shiota, K (2007) Genome-wide and locus-specific DNA hypomethylation in G9a deficient mouse embryonic stem cells. Genes Cells 12:1-11

Abstract

In the mammalian genome, numerous CpG-rich loci define tissue-dependent and differentially methylated regions (T-DMRs). Euchromatin from different cell types differs in terms of its tissue-specific DNA methylation profile as defined by these T-DMRs. G9a is a euchromatin-localized histone methyltransferase (HMT) and catalyzes methylation of histone H3 at lysines 9 and 27 (H3-K9 and -K27). To test whether HMT activity influences euchromatic cytosine methylation, we analyzed the DNA methylation status of approximately 2000 CpG-rich loci, which are predicted in silico, in G9a(-/-) embryonic stem cells by restriction landmark genomic scanning (RLGS). While the RLGS profile of wild-type cells contained about 1300 spots, 32 new spots indicating DNA demethylation were seen in the profile of G9a(-/-) cells. Virtual-image RLGS (Vi-RLGS) allowed us to identify the genomic source of ten of these spots. These were confirmed to be cytosine demethylated, not just at the Not I site detected by the RLGS but extending over several kilobase pairs in cis. Chromatin immunoprecipitation (ChIP) confirmed these loci to be targets of G9a, with decreased H3-K9 and/or -K27 dimethylation in the G9a(-/-) cells. These data indicate that G9a site-selectively contributes to DNA methylation.

Links

PubMed Online version:10.1111/j.1365-2443.2006.01029.x

Keywords

Animals; Chromatin Immunoprecipitation; CpG Islands; Cysteine/metabolism; DNA/metabolism; DNA Methylation; Embryonic Stem Cells/enzymology; Euchromatin/enzymology; Euchromatin/metabolism; Gene Deletion; Genome; Histone-Lysine N-Methyltransferase/genetics; Histone-Lysine N-Methyltransferase/metabolism; Histones/metabolism; Mice; Models, Genetic; Nucleosomes/metabolism; Polymerase Chain Reaction; Protein Methyltransferases

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:EHMT2

acts_upstream_of_or_within

GO:0010424: DNA methylation on cytosine within a CG sequence

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:EHMT2

acts_upstream_of_or_within

GO:0051567: histone H3-K9 methylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:EHMT2

acts_upstream_of_or_within

GO:0070734: histone H3-K27 methylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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