Inhibition of DNA methyltransferase leads to increased genomic 5-hydroxymethylcytosine levels in hematopoietic cells

Borbála Vető, P. Szabó, Caroline Bacquet, Anna Apró, Edit Hathy, Judit Kiss, J. Réthelyi, Flóra Szeri, D. Szüts, T. Arányi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

5-Hydroxymethylcytosine (5hmC) is produced from 5-methylcytosine (5mC) by Ten-eleven translocation (TET) dioxygenases. The epigenetic modification 5hmC has crucial roles in both cellular development and differentiation. The 5hmC level is particularly high in the brain. While 5mC is generally associated with gene silencing/reduced expression, 5hmC is a more permissive epigenetic mark. To understand its physiological function, an easy and accurate quantification method is required. Here, we have developed a novel LC-MS/MS-based approach to quantify both genomic 5mC and 5hmC contents. The method is based on the liberation of nucleobases by formic acid. Applying this method, we characterized the levels of DNA methylation and hydroxymethylation in mouse brain and liver, primary hepatocytes, and various cell lines. Using this approach, we confirm that the treatment of different cell lines with the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine leads to a decrease in 5mC content. This decrease was accompanied by an increase in 5hmC levels in cell lines of hematopoietic origin. Finally, we showed that ascorbate elevates the levels of 5hmC and augments the effect of 5-aza-2′-deoxycytidine without significantly influencing 5mC levels.

Original languageEnglish
Pages (from-to)584-592
Number of pages9
JournalFEBS Open Bio
Volume8
Issue number4
DOIs
Publication statusPublished - Apr 1 2018

Fingerprint

Methyltransferases
5-Methylcytosine
decitabine
DNA
formic acid
Cells
Epigenomics
Cell Line
Brain
Dioxygenases
Gene Silencing
DNA Methylation
5-hydroxymethylcytosine
Liver
Hepatocytes
Genes

Keywords

  • 5-aza-2′-deoxycytidine
  • 5-hydroxymethylcytosine
  • 5-methylcytosine
  • ascorbate
  • LC-MS/MS

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Inhibition of DNA methyltransferase leads to increased genomic 5-hydroxymethylcytosine levels in hematopoietic cells. / Vető, Borbála; Szabó, P.; Bacquet, Caroline; Apró, Anna; Hathy, Edit; Kiss, Judit; Réthelyi, J.; Szeri, Flóra; Szüts, D.; Arányi, T.

In: FEBS Open Bio, Vol. 8, No. 4, 01.04.2018, p. 584-592.

Research output: Contribution to journalArticle

Vető, Borbála ; Szabó, P. ; Bacquet, Caroline ; Apró, Anna ; Hathy, Edit ; Kiss, Judit ; Réthelyi, J. ; Szeri, Flóra ; Szüts, D. ; Arányi, T. / Inhibition of DNA methyltransferase leads to increased genomic 5-hydroxymethylcytosine levels in hematopoietic cells. In: FEBS Open Bio. 2018 ; Vol. 8, No. 4. pp. 584-592.
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