TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation

TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation

The discovery of hydroxymethyl-, formyl- and carboxylcytosine, generated through oxidation of methylcytosine by TET dioxygenases, raised the question how these modifications contribute to epigenetic regulation. As they are subjected to complex regulation in vivo, we dissected links to gene expressio...

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Main Authors: Müller, Udo, Bauer, Christina, Siegl, Michael, Rottach, Andrea, Leonhardt, Heinrich
Format: Online
Language:English
Published: Oxford University Press 2014
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4117777/
id pubmed-4117777
recordtype oai_dc
spelling pubmed-41177772014-08-15 TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation Müller, Udo Bauer, Christina Siegl, Michael Rottach, Andrea Leonhardt, Heinrich Gene regulation, Chromatin and Epigenetics The discovery of hydroxymethyl-, formyl- and carboxylcytosine, generated through oxidation of methylcytosine by TET dioxygenases, raised the question how these modifications contribute to epigenetic regulation. As they are subjected to complex regulation in vivo, we dissected links to gene expression with in vitro modified reporter constructs. We used an Oct4 promoter-driven reporter gene and demonstrated that in vitro methylation causes gene silencing while subsequent oxidation with purified catalytic domain of TET1 leads to gene reactivation. To identify proteins involved in this pathway we screened for TET interacting factors and identified TDG, PARP1, XRCC1 and LIG3 that are involved in base-excision repair. Knockout and rescue experiments demonstrated that gene reactivation depended on the glycosylase TDG, but not MBD4, while NEIL1, 2 and 3 could partially rescue the loss of TDG. These results clearly show that oxidation of methylcytosine by TET dioxygenases and subsequent removal by TDG or NEIL glycosylases and the BER pathway results in reactivation of epigenetically silenced genes. Oxford University Press 2014-09-01 2014-06-21 /pmc/articles/PMC4117777/ /pubmed/24948610 http://dx.doi.org/10.1093/nar/gku552 Text en © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Müller, Udo
Bauer, Christina
Siegl, Michael
Rottach, Andrea
Leonhardt, Heinrich
spellingShingle Müller, Udo
Bauer, Christina
Siegl, Michael
Rottach, Andrea
Leonhardt, Heinrich
TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation
author_facet Müller, Udo
Bauer, Christina
Siegl, Michael
Rottach, Andrea
Leonhardt, Heinrich
author_sort Müller, Udo
title TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation
title_short TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation
title_full TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation
title_fullStr TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation
title_full_unstemmed TET-mediated oxidation of methylcytosine causes TDG or NEIL glycosylase dependent gene reactivation
title_sort tet-mediated oxidation of methylcytosine causes tdg or neil glycosylase dependent gene reactivation
description The discovery of hydroxymethyl-, formyl- and carboxylcytosine, generated through oxidation of methylcytosine by TET dioxygenases, raised the question how these modifications contribute to epigenetic regulation. As they are subjected to complex regulation in vivo, we dissected links to gene expression with in vitro modified reporter constructs. We used an Oct4 promoter-driven reporter gene and demonstrated that in vitro methylation causes gene silencing while subsequent oxidation with purified catalytic domain of TET1 leads to gene reactivation. To identify proteins involved in this pathway we screened for TET interacting factors and identified TDG, PARP1, XRCC1 and LIG3 that are involved in base-excision repair. Knockout and rescue experiments demonstrated that gene reactivation depended on the glycosylase TDG, but not MBD4, while NEIL1, 2 and 3 could partially rescue the loss of TDG. These results clearly show that oxidation of methylcytosine by TET dioxygenases and subsequent removal by TDG or NEIL glycosylases and the BER pathway results in reactivation of epigenetically silenced genes.
publisher Oxford University Press
publishDate 2014
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4117777/
_version_ 1613119645496115200

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