Acute hydroxyurea-induced replication blockade results in replisome components disengagement from nascent DNA without causing fork collapse

Amaia Ercilla; Sonia Feu; Sergi Aranda; Alba Llopis; Sólveig Hlín Brynjólfsdóttir; Claus Storgaard Sørensen; Luis Ignacio Toledo; Neus Agell

doi:10.1007/s00018-019-03206-1

Acute hydroxyurea-induced replication blockade results in replisome components disengagement from nascent DNA without causing fork collapse

Amaia Ercilla, Sonia Feu, Sergi Aranda, Alba Llopis, Sólveig Hlín Brynjólfsdóttir, Claus Storgaard Sørensen, Luis Ignacio Toledo, Neus Agell

Molecular Aging Program

5 Citations (Scopus)

Abstract

During S phase, replication forks can encounter several obstacles that lead to fork stalling, which if persistent might result in fork collapse. To avoid this collapse and to preserve the competence to restart, cells have developed mechanisms that maintain fork stability upon replication stress. In this study, we aimed to understand the mechanisms involved in fork stability maintenance in non-transformed human cells by performing an isolation of proteins on nascent DNA-mass spectrometry analysis in hTERT-RPE cells under different replication stress conditions. Our results show that acute hydroxyurea-induced replication blockade causes the accumulation of large amounts of single-stranded DNA at the fork. Remarkably, this results in the disengagement of replisome components from nascent DNA without compromising fork restart. Notably, Cdc45-MCM-GINS helicase maintains its integrity and replisome components remain associated with chromatin upon acute hydroxyurea treatment, whereas replisome stability is lost upon a sustained replication stress that compromises the competence to restart.

Original language	English
Journal	Cellular and molecular life sciences : CMLS
ISSN	1420-682X
DOIs	https://doi.org/10.1007/s00018-019-03206-1
Publication status	Published - 1 Feb 2020

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title = "Acute hydroxyurea-induced replication blockade results in replisome components disengagement from nascent DNA without causing fork collapse",

abstract = "During S phase, replication forks can encounter several obstacles that lead to fork stalling, which if persistent might result in fork collapse. To avoid this collapse and to preserve the competence to restart, cells have developed mechanisms that maintain fork stability upon replication stress. In this study, we aimed to understand the mechanisms involved in fork stability maintenance in non-transformed human cells by performing an isolation of proteins on nascent DNA-mass spectrometry analysis in hTERT-RPE cells under different replication stress conditions. Our results show that acute hydroxyurea-induced replication blockade causes the accumulation of large amounts of single-stranded DNA at the fork. Remarkably, this results in the disengagement of replisome components from nascent DNA without compromising fork restart. Notably, Cdc45-MCM-GINS helicase maintains its integrity and replisome components remain associated with chromatin upon acute hydroxyurea treatment, whereas replisome stability is lost upon a sustained replication stress that compromises the competence to restart.",

author = "Amaia Ercilla and Sonia Feu and Sergi Aranda and Alba Llopis and Brynj{\'o}lfsd{\'o}ttir, {S{\'o}lveig Hl{\'i}n} and S{\o}rensen, {Claus Storgaard} and Toledo, {Luis Ignacio} and Neus Agell",

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doi = "10.1007/s00018-019-03206-1",

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T1 - Acute hydroxyurea-induced replication blockade results in replisome components disengagement from nascent DNA without causing fork collapse

AU - Ercilla, Amaia

AU - Feu, Sonia

AU - Aranda, Sergi

AU - Llopis, Alba

AU - Brynjólfsdóttir, Sólveig Hlín

AU - Sørensen, Claus Storgaard

AU - Toledo, Luis Ignacio

AU - Agell, Neus

PY - 2020/2/1

Y1 - 2020/2/1

N2 - During S phase, replication forks can encounter several obstacles that lead to fork stalling, which if persistent might result in fork collapse. To avoid this collapse and to preserve the competence to restart, cells have developed mechanisms that maintain fork stability upon replication stress. In this study, we aimed to understand the mechanisms involved in fork stability maintenance in non-transformed human cells by performing an isolation of proteins on nascent DNA-mass spectrometry analysis in hTERT-RPE cells under different replication stress conditions. Our results show that acute hydroxyurea-induced replication blockade causes the accumulation of large amounts of single-stranded DNA at the fork. Remarkably, this results in the disengagement of replisome components from nascent DNA without compromising fork restart. Notably, Cdc45-MCM-GINS helicase maintains its integrity and replisome components remain associated with chromatin upon acute hydroxyurea treatment, whereas replisome stability is lost upon a sustained replication stress that compromises the competence to restart.

AB - During S phase, replication forks can encounter several obstacles that lead to fork stalling, which if persistent might result in fork collapse. To avoid this collapse and to preserve the competence to restart, cells have developed mechanisms that maintain fork stability upon replication stress. In this study, we aimed to understand the mechanisms involved in fork stability maintenance in non-transformed human cells by performing an isolation of proteins on nascent DNA-mass spectrometry analysis in hTERT-RPE cells under different replication stress conditions. Our results show that acute hydroxyurea-induced replication blockade causes the accumulation of large amounts of single-stranded DNA at the fork. Remarkably, this results in the disengagement of replisome components from nascent DNA without compromising fork restart. Notably, Cdc45-MCM-GINS helicase maintains its integrity and replisome components remain associated with chromatin upon acute hydroxyurea treatment, whereas replisome stability is lost upon a sustained replication stress that compromises the competence to restart.

U2 - 10.1007/s00018-019-03206-1

DO - 10.1007/s00018-019-03206-1

M3 - Journal article

C2 - 31297568

SN - 1420-682X

JO - Cellular and molecular life sciences : CMLS

JF - Cellular and molecular life sciences : CMLS

ER -

Acute hydroxyurea-induced replication blockade results in replisome components disengagement from nascent DNA without causing fork collapse

Abstract

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