Sensitive enzymatic quantification of 5-hydroxymethylcytosine in genomic DNA.
The recent discovery of genomic 5-hydroxymethylcytosine (hmC) and mutations affecting the respective Tet hydroxylases in leukemia raises fundamental questions about this epigenetic modification. We present a sensitive method for fast quantification of genomic hmC based on specific transfer of radiolabeled glucose to hmC by a purified glucosyltransferase. We determined hmC levels in various adult tissues and differentiating embryonic stem cells and show a correlation with differential expression of tet genes.
Szwagierczak
Aleksandra
A
Department of Biology, Center for Integrated Protein Science Munich, Ludwig Maximilians University Munich, 82152 Planegg-Martinsried, Germany.
Bultmann
Sebastian
S
Schmidt
Christine S
CS
Spada
Fabio
F
Leonhardt
Heinrich
H
D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000818:Animals; D002478:Cells, Cultured; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000737:chemistry / Q000378:metabolism; D005821:Genetic Techniques; D016678:Genome; D005964:Glucosyltransferases / Q000378:metabolism; D051379:Mice
C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D005964:Glucosyltransferases