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dc.contributor.authorMorafraile, Esther C.
dc.contributor.authorBugallo, Alberto
dc.contributor.authorCarreira Rodríguez, Raquel
dc.contributor.authorFernández, María
dc.contributor.authorMartín Castellanos, Cristina
dc.contributor.authorGonzález Blanco, Miguel
dc.contributor.authorSegurado, Mónica
dc.date.accessioned2021-02-16T13:28:19Z
dc.date.available2021-02-16T13:28:19Z
dc.date.issued2020
dc.identifier.citationNucleic Acids Research, Volume 48, Issue 6, 06 April 2020, Pages 3053–3070, https://doi.org/10.1093/nar/gkaa054
dc.identifier.issn0305-1048
dc.identifier.urihttp://hdl.handle.net/10347/24486
dc.description.abstractThe S phase checkpoint is crucial to maintain genome stability under conditions that threaten DNA replication. One of its critical functions is to prevent Exo1-dependent fork degradation, and Exo1 is phosphorylated in response to different genotoxic agents. Exo1 seemed to be regulated by several post-translational modifications in the presence of replicative stress, but the specific contribution of checkpoint-dependent phosphorylation to Exo1 control and fork stability is not clear. We show here that Exo1 phosphorylation is Dun1-independent and Rad53-dependent in response to DNA damage or dNTP depletion, and in both situations Exo1 is similarly phosphorylated at multiple sites. To investigate the correlation between Exo1 phosphorylation and fork stability, we have generated phospho-mimic exo1 alleles that rescue fork collapse in rad53 mutants as efficiently as exo1-nuclease dead mutants or the absence of Exo1, arguing that Rad53-dependent phosphorylation is the mayor requirement to preserve fork stability. We have also shown that this rescue is Bmh1–2 independent, arguing that the 14-3-3 proteins are dispensable for fork stabilization, at least when Exo1 is downregulated. Importantly, our results indicated that phosphorylation specifically inhibits the 5' to 3'exo-nuclease activity, suggesting that this activity of Exo1 and not the flap-endonuclease, is the enzymatic activity responsible of the collapse of stalled replication forks in checkpoint mutants
dc.description.sponsorshipSpanish Ministry of Economy and Competitiveness (MINECO) [FEDER-BFU2013-45182-P to C.M.C., M.S.]; University of Salamanca [KA6H/463AC01 to M.S.]; MINECO, AEI, Xunta de Galicia and FEDER [RYC-2012-10835, BFU2016-78121-P, ED431F-2016/019, ED431B-2016/016 to M.G.B.]; Junta de Castilla y León (JCyL), Program ‘Escalera de Excelencia’ [FEDER-CLU-2017-03]; JCyL Pre-doctoral Fellowship (to A.B.); MINECO Pre-Doctoral Fellowship (to E.C.M.); Xunta de Galicia Pre-doctoral Fellowship (ED481A-2018/041 to R.C.)
dc.language.isoeng
dc.publisherOxford University Press
dc.relationinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2013-45182-P/ES/REGULACION DE LA INTEGRIDAD DEL GENOMA: REPLICACION Y RECOMBINACION
dc.rights© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), 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
dc.rightsAtribución-NoComercial 4.0 Internacional
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/
dc.titleExo1 phosphorylation inhibits exonuclease activity and prevents fork collapse in rad53 mutants independently of the 14-3-3 proteins
dc.typeinfo:eu-repo/semantics/article
dc.identifier.DOI10.1093/nar/gkaa054
dc.relation.publisherversionhttps://doi.org/10.1093/nar/gkaa054
dc.type.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.e-issn1362-4962
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.contributor.affiliationUniversidade de Santiago de Compostela. Centro de Investigación en Medicina Molecular e Enfermidades Crónicas
dc.contributor.affiliationUniversidade de Santiago de Compostela. Departamento de Bioquímica e Bioloxía Molecular
dc.description.peerreviewedSI


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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), 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
Except where otherwise noted, this item's license is described as  © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), 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





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