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Author  |
Kinney, S.M.; Chin, H.G.; Vaisvila, R.; Bitinaite, J.; Zheng, Y.; Esteve, P.O.; Feng, S.; Stroud, H.; Jacobsen, S.E.; Pradhan, S. |
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Title |
Tissue specific distribution and dynamic changes of 5-hydroxymethylcytosine in mammalian genome |
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Journal Article |
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Year |
2011 |
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The Journal of Biological Chemistry |
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J Biol Chem |
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Abstract |
Cytosine residues in the vertebrate genome are enzymatically modified to 5-methylcytosine, which participates in transcriptional repression of genes during development and disease progression. 5-methylcytosine can be further enzymatically modified to 5-hydroxymethylcytosine by the TET family of methylcytosine dioxygenases. Analysis of 5-methylcytosine and 5-hydroxymethylcytosine is confounded, as these modifications are indistinguishable by traditional sequencing methods even when supplemented by bisulfite conversion. Here we demonstrate a simple enzymatic approach that involves cloning, identification, and quantification of 5-hydroxymethylcytosine in various CCGG loci within murine and human genomes. 5-hydroxymethylcytosine was prevalent in human and murine brain and heart genomic DNAs at several regions. The cultured cell lines NIH3T3 and HeLa both displayed very low or undetectable amounts of 5-hydroxymethylcytosine at the examined loci. Interestingly, 5-hydroxymethylcytosine levels in mouse embryonic stem cell DNA first increased then slowly decreased upon differentiation to embryoid bodies whereas 5-methylcytosine levels increased gradually over time. Finally, using a quantitative PCR approach, we established that a portion of VANGL1 and EGFR gene body methylation in human tissue DNA samples is indeed hydroxymethylation. |
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New England Biolabs, United States |
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English |
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0021-9258 |
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Notes |
PMID:21610077 |
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Call Number |
pubs @ mcanady @ |
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23488 |
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