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Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells
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 associa...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5881552/ https://www.ncbi.nlm.nih.gov/pubmed/29632811 http://dx.doi.org/10.1002/2211-5463.12392 |
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author | Vető, Borbála Szabó, Pál Bacquet, Caroline Apró, Anna Hathy, Edit Kiss, Judit Réthelyi, János M. Szeri, Flóra Szüts, Dávid Arányi, Tamás |
author_facet | Vető, Borbála Szabó, Pál Bacquet, Caroline Apró, Anna Hathy, Edit Kiss, Judit Réthelyi, János M. Szeri, Flóra Szüts, Dávid Arányi, Tamás |
author_sort | Vető, Borbála |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5881552 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-58815522018-04-09 Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells Vető, Borbála Szabó, Pál Bacquet, Caroline Apró, Anna Hathy, Edit Kiss, Judit Réthelyi, János M. Szeri, Flóra Szüts, Dávid Arányi, Tamás FEBS Open Bio Research Articles 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. John Wiley and Sons Inc. 2018-02-23 /pmc/articles/PMC5881552/ /pubmed/29632811 http://dx.doi.org/10.1002/2211-5463.12392 Text en © 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Vető, Borbála Szabó, Pál Bacquet, Caroline Apró, Anna Hathy, Edit Kiss, Judit Réthelyi, János M. Szeri, Flóra Szüts, Dávid Arányi, Tamás Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
title | Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
title_full | Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
title_fullStr | Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
title_full_unstemmed | Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
title_short | Inhibition of DNA methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
title_sort | inhibition of dna methyltransferase leads to increased genomic 5‐hydroxymethylcytosine levels in hematopoietic cells |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5881552/ https://www.ncbi.nlm.nih.gov/pubmed/29632811 http://dx.doi.org/10.1002/2211-5463.12392 |
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