A Short G1 Phase Imposes Constitutive Replication Stress and Fork Remodelling in Mouse Embryonic Stem Cells

Overview

Journal Nat Commun

Specialty Biology

Date 2016 Feb 16

PMID 26876348

Citations 103

Authors

Akshay K Ahuja

Karolina Jodkowska

Federico Teloni

Anna H Bizard

Ralph Zellweger

Raquel Herrador

Sagrario Ortega

Ian D Hickson

Matthias Altmeyer

Juan Mendez

Massimo Lopes

Affiliations

Soon will be listed here.

Abstract

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.

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