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

Luis J. García-Rodríguez, Giacomo De Piccoli, Vanessa Marchesi, Richard C. Jones, Ricky D. Edmondson, Karim Labib (Lead / Corresponding author)

Research output: Contribution to journalArticle

27 Citations (Scopus)
144 Downloads (Pure)

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.
Original languageEnglish
Pages (from-to)8830-8838
JournalNucleic Acids Research
Volume43
Issue number18
Early online date6 Aug 2015
DOIs
Publication statusPublished - 15 Oct 2015

Fingerprint Dive into the research topics of 'A conserved Polϵ binding module in Ctf18-RFC is required for S-phase checkpoint activation downstream of Mec1'. Together they form a unique fingerprint.

  • Projects

    Profiles

    No photo of Karim Labib

    Labib, Karim

    • MRC PPU - Professor & Personal Chair Genome Integrity

    Person: Academic

    Cite this

    García-Rodríguez, L. J., De Piccoli, G., Marchesi, V., Jones, R. C., Edmondson, R. D., & Labib, K. (2015). A conserved Polϵ binding module in Ctf18-RFC is required for S-phase checkpoint activation downstream of Mec1. Nucleic Acids Research, 43(18), 8830-8838. https://doi.org/10.1093/nar/gkv799