Format

Send to

Choose Destination
PLoS One. 2010 Dec 23;5(12):e15367. doi: 10.1371/journal.pone.0015367.

Tissue distribution of 5-hydroxymethylcytosine and search for active demethylation intermediates.

Author information

1
Department of Chemistry, Ludwig-Maximilians-University, Center for Integrated Protein Science, Munich, Germany.

Abstract

5-Hydroxymethylcytosine (hmC) was recently detected as the sixth base in mammalian tissue at so far controversial levels. The function of the modified base is currently unknown, but it is certain that the base is generated from 5-methylcytosine (mC). This fuels the hypothesis that it represents an intermediate of an active demethylation process, which could involve further oxidation of the hydroxymethyl group to a formyl or carboxyl group followed by either deformylation or decarboxylation. Here, we use an ultra-sensitive and accurate isotope based LC-MS method to precisely determine the levels of hmC in various mouse tissues and we searched for 5-formylcytosine (fC), 5-carboxylcytosine (caC), and 5-hydroxymethyluracil (hmU) as putative active demethylation intermediates. Our data suggest that an active oxidative mC demethylation pathway is unlikely to occur. Additionally, we show using HPLC-MS analysis and immunohistochemistry that hmC is present in all tissues and cell types with highest concentrations in neuronal cells of the CNS.

PMID:
21203455
PMCID:
PMC3009720
DOI:
10.1371/journal.pone.0015367
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Public Library of Science Icon for PubMed Central
Loading ...
Support Center