Sensitive enzymatic quantification of 5-hydroxymethylcytosine in genomic DNA

Nucleic Acids Res. 2010 Oct;38(19):e181. doi: 10.1093/nar/gkq684. Epub 2010 Aug 4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5-Methylcytosine / analogs & derivatives
  • Animals
  • Cells, Cultured
  • Cytosine / analogs & derivatives*
  • Cytosine / analysis
  • Cytosine / metabolism
  • DNA / chemistry*
  • Embryonic Stem Cells / chemistry
  • Embryonic Stem Cells / metabolism
  • Genetic Techniques
  • Genome
  • Glucosyltransferases / metabolism
  • Mice

Substances

  • 5-hydroxymethylcytosine
  • 5-Methylcytosine
  • Cytosine
  • DNA
  • Glucosyltransferases