A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells
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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.
I: Nature Communications, Bind 7, 10660, 02.2016.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells
AU - Ahuja, Akshay K.
AU - Jodkowska, Karolina
AU - Teloni, Federico
AU - Bizard, Anna H.
AU - Zellweger, Ralph
AU - Herrador, Raquel
AU - Ortega, Sagrario
AU - Hickson, Ian D.
AU - Altmeyer, Matthias
AU - Mendez, Juan
AU - Lopes, Massimo
PY - 2016/2
Y1 - 2016/2
N2 - 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.
AB - 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.
U2 - 10.1038/ncomms10660
DO - 10.1038/ncomms10660
M3 - Journal article
C2 - 26876348
VL - 7
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
M1 - 10660
ER -
ID: 167480205