Publication Date:
2012-04-28
Description:
5-Methylcytosine can be converted to 5-hydroxymethylcytosine (5hmC) in mammalian DNA by the ten-eleven translocation (TET) enzymes. We introduce oxidative bisulfite sequencing (oxBS-Seq), the first method for quantitative mapping of 5hmC in genomic DNA at single-nucleotide resolution. Selective chemical oxidation of 5hmC to 5-formylcytosine (5fC) enables bisulfite conversion of 5fC to uracil. We demonstrate the utility of oxBS-Seq to map and quantify 5hmC at CpG islands (CGIs) in mouse embryonic stem (ES) cells and identify 800 5hmC-containing CGIs that have on average 3.3% hydroxymethylation. High levels of 5hmC were found in CGIs associated with transcriptional regulators and in long interspersed nuclear elements, suggesting that these regions might undergo epigenetic reprogramming in ES cells. Our results open new questions on 5hmC dynamics and sequence-specific targeting by TETs.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Booth, Michael J -- Branco, Miguel R -- Ficz, Gabriella -- Oxley, David -- Krueger, Felix -- Reik, Wolf -- Balasubramanian, Shankar -- 095645/Wellcome Trust/United Kingdom -- 11961/Cancer Research UK/United Kingdom -- G0801156/Medical Research Council/United Kingdom -- Biotechnology and Biological Sciences Research Council/United Kingdom -- Cancer Research UK/United Kingdom -- Medical Research Council/United Kingdom -- Wellcome Trust/United Kingdom -- New York, N.Y. -- Science. 2012 May 18;336(6083):934-7. doi: 10.1126/science.1220671. Epub 2012 Apr 26.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/22539555" target="_blank"〉PubMed〈/a〉
Keywords:
5-Methylcytosine/*analysis
;
Animals
;
*CpG Islands
;
Cytosine/*analogs & derivatives/analysis/chemistry
;
DNA/*chemistry/genetics
;
DNA Methylation
;
*Embryonic Stem Cells/physiology
;
Epigenesis, Genetic
;
Genes, Intracisternal A-Particle
;
High-Throughput Nucleotide Sequencing
;
Long Interspersed Nucleotide Elements
;
Mice
;
Oxidation-Reduction
;
Rhenium/chemistry
;
*Sequence Analysis, DNA
;
Sulfites
;
Transcription, Genetic
;
Uracil/chemistry
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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