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PMID: 21203455 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

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

PloS one ·Vol. 5 ·No. 12 ·2010-12-23 ·Pages e15367

Globisch D, Münzel M, Müller M, Michalakis S, Wagner M, Koch S, Brückl T, Biel M, Carell T

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.

MeSH Terms
5-Methylcytosine/analogs & derivatives Animals Calibration Chromatography, High Pressure Liquid/methods Chromatography, Liquid/methods Cytosine/analogs & derivatives,chemistry,metabolism DNA Methylation Immunohistochemistry/methods Mass Spectrometry/methods Mice Models, Chemical Oxygen/chemistry Spectrometry, Mass, Electrospray Ionization/methods Tissue Distribution
Chemicals
5-hydroxymethylcytosine 5-Methylcytosine Cytosine Oxygen
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Globisch Daniel
Department of Chemistry, Ludwig-Maximilians-University, Center for Integrated Protein Science, Munich, Germany.
Münzel Martin
Müller Markus
Michalakis Stylianos
Wagner Mirko
Koch Susanne
Brückl Tobias
Biel Martin
Carell Thomas
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Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0015367
Published
2010-12-23
Epub
2010-00-23
Pages
e15367
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC3009720
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0015367 PMC Full Text View journal details
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