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    A conserved Polϵ binding module in Ctf18-RFC is required for S-phase checkpoint activation downstream of Mec1.

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    Authors
    García-Rodríguez, Luis J
    De Piccoli, G
    Marchesi, Vanessa
    Jones, R
    Edmondson, R
    Labib, K
    Affiliation
    Cancer Research UK Manchester Institute, University of Manchester, Wilmslow Road, Manchester M20 4BX
    Issue Date
    2015-08-06
    
    Metadata
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    Abstract
    Defects during chromosome replication in eukaryotes activate a signaling pathway called the S-phase checkpoint, which produces a multifaceted response that preserves genome integrity at stalled DNA replication forks. Work with budding yeast showed that the 'alternative clamp loader' known as Ctf18-RFC acts by an unknown mechanism to activate the checkpoint kinase Rad53, which then mediates much of the checkpoint response. Here we show that budding yeast Ctf18-RFC associates with DNA polymerase epsilon, via an evolutionarily conserved 'Pol ϵ binding module' in Ctf18-RFC that is produced by interaction of the carboxyl terminus of Ctf18 with the Ctf8 and Dcc1 subunits. Mutations at the end of Ctf18 disrupt the integrity of the Pol ϵ binding module and block the S-phase checkpoint pathway, downstream of the Mec1 kinase that is the budding yeast orthologue of mammalian ATR. Similar defects in checkpoint activation are produced by mutations that displace Pol ϵ from the replisome. These findings indicate that the association of Ctf18-RFC with Pol ϵ at defective replication forks is a key step in activation of the S-phase checkpoint.
    Citation
    A conserved Pol binding module in Ctf18-RFC is required for S-phase checkpoint activation downstream of Mec1. 2015: Nucleic Acids Res
    Journal
    Nucleic Acids Research
    URI
    http://hdl.handle.net/10541/581078
    DOI
    10.1093/nar/gkv799
    PubMed ID
    26250113
    Type
    Article
    Language
    en
    ISSN
    1362-4962
    ae974a485f413a2113503eed53cd6c53
    10.1093/nar/gkv799
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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