期刊
EPIGENETICS
卷 7, 期 2, 页码 131-136出版社
LANDES BIOSCIENCE
DOI: 10.4161/epi.7.2.18909
关键词
hydroxymethylcytosine; MspI; epigenetics; DNA methylation; endonuclease
资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Grants-in-Aid for Scientific Research [22770009, 21200037] Funding Source: KAKEN
In mammalian genomic DNA, cytosine methylation predominantly occurs at CpG dinucleotides and provides epigenetic information. In some cells, 5-methyl-cytosine (5-mC) can be further converted to 5-hydroxymethyl-cytosine (5-hmC) by the ten-eleven translocation family of proteins. MspI restriction endonuclease has been used to analyze these modified cytosines. However, the kinetic analysis in this study revealed that MspI activity is dramatically decreased by symmetrical hydroxymethylation of its recognition sequence and partly inhibited by hemi-hydroxymethylation, whereas TaqI and HaeIII are relatively resistant to hydroxymethylation. Therefore, DNA modification studies that use MspI, for example, reduced representation bisulfite shotgun sequencing, quantitative analysis of 5-hmC and cleavage-sensitivity analysis, should be carefully interpreted.
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