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A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells
Embryonic stem cells (ESCs) represent a transient biological state, where pluripotency is coupled with fast proliferation. ESCs display a constitutively active DNA damage response (DDR), but its molecular determinants have remained elusive. Here we show in cultured ESCs and mouse embryos that H2AX p...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4756311/ https://www.ncbi.nlm.nih.gov/pubmed/26876348 http://dx.doi.org/10.1038/ncomms10660 |
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author | Ahuja, Akshay K. Jodkowska, Karolina Teloni, Federico Bizard, Anna H. Zellweger, Ralph Herrador, Raquel Ortega, Sagrario Hickson, Ian D. Altmeyer, Matthias Mendez, Juan Lopes, Massimo |
author_facet | Ahuja, Akshay K. Jodkowska, Karolina Teloni, Federico Bizard, Anna H. Zellweger, Ralph Herrador, Raquel Ortega, Sagrario Hickson, Ian D. Altmeyer, Matthias Mendez, Juan Lopes, Massimo |
author_sort | Ahuja, Akshay K. |
collection | PubMed |
description | Embryonic stem cells (ESCs) represent a transient biological state, where pluripotency is coupled with fast proliferation. ESCs display a constitutively active DNA damage response (DDR), but its molecular determinants have remained elusive. Here we show in cultured ESCs and mouse embryos that H2AX phosphorylation is dependent on Ataxia telangiectasia and Rad3 related (ATR) and is associated with chromatin loading of the ssDNA-binding proteins RPA and RAD51. Single-molecule analysis of replication intermediates reveals massive ssDNA gap accumulation, reduced fork speed and frequent fork reversal. All these marks of replication stress do not impair the mitotic process and are rapidly lost at differentiation onset. Delaying the G1/S transition in ESCs allows formation of 53BP1 nuclear bodies and suppresses ssDNA accumulation, fork slowing and reversal in the following S-phase. Genetic inactivation of fork slowing and reversal leads to chromosomal breakage in unperturbed ESCs. We propose that rapid cell cycle progression makes ESCs dependent on effective replication-coupled mechanisms to protect genome integrity. |
format | Online Article Text |
id | pubmed-4756311 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-47563112016-03-04 A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells Ahuja, Akshay K. Jodkowska, Karolina Teloni, Federico Bizard, Anna H. Zellweger, Ralph Herrador, Raquel Ortega, Sagrario Hickson, Ian D. Altmeyer, Matthias Mendez, Juan Lopes, Massimo Nat Commun Article Embryonic stem cells (ESCs) represent a transient biological state, where pluripotency is coupled with fast proliferation. ESCs display a constitutively active DNA damage response (DDR), but its molecular determinants have remained elusive. Here we show in cultured ESCs and mouse embryos that H2AX phosphorylation is dependent on Ataxia telangiectasia and Rad3 related (ATR) and is associated with chromatin loading of the ssDNA-binding proteins RPA and RAD51. Single-molecule analysis of replication intermediates reveals massive ssDNA gap accumulation, reduced fork speed and frequent fork reversal. All these marks of replication stress do not impair the mitotic process and are rapidly lost at differentiation onset. Delaying the G1/S transition in ESCs allows formation of 53BP1 nuclear bodies and suppresses ssDNA accumulation, fork slowing and reversal in the following S-phase. Genetic inactivation of fork slowing and reversal leads to chromosomal breakage in unperturbed ESCs. We propose that rapid cell cycle progression makes ESCs dependent on effective replication-coupled mechanisms to protect genome integrity. Nature Publishing Group 2016-02-15 /pmc/articles/PMC4756311/ /pubmed/26876348 http://dx.doi.org/10.1038/ncomms10660 Text en Copyright © 2016, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Ahuja, Akshay K. Jodkowska, Karolina Teloni, Federico Bizard, Anna H. Zellweger, Ralph Herrador, Raquel Ortega, Sagrario Hickson, Ian D. Altmeyer, Matthias Mendez, Juan Lopes, Massimo A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
title | A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
title_full | A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
title_fullStr | A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
title_full_unstemmed | A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
title_short | A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
title_sort | short g1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4756311/ https://www.ncbi.nlm.nih.gov/pubmed/26876348 http://dx.doi.org/10.1038/ncomms10660 |
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