Analysis of replication profiles reveals key role of RFC-Ctf18 in yeast replication stress response

Laure Crabbé; Aubin Thomas; Véronique Pantesco; John De Vos; Philippe Pasero; Armelle Lengronne

doi:10.1038/nsmb.1932

Analysis of replication profiles reveals key role of RFC-Ctf18 in yeast replication stress response

Nat Struct Mol Biol. 2010 Nov;17(11):1391-7. doi: 10.1038/nsmb.1932. Epub 2010 Oct 24.

Authors

Laure Crabbé¹, Aubin Thomas, Véronique Pantesco, John De Vos, Philippe Pasero, Armelle Lengronne

Affiliation

¹ Institute of Human Genetics, Centre National de Recherche Scientifique, Unité Propre de Recherche 1142, Montpellier, France.

PMID: 20972444
DOI: 10.1038/nsmb.1932

Abstract

Maintenance of genome integrity relies on surveillance mechanisms that detect and signal arrested replication forks. Although evidence from budding yeast indicates that the DNA replication checkpoint (DRC) is primarily activated by single-stranded DNA (ssDNA), studies in higher eukaryotes have implicated primer ends in this process. To identify factors that signal primed ssDNA in Saccharomyces cerevisiae, we have screened a collection of checkpoint mutants for their ability to activate the DRC, using the repression of late origins as readout for checkpoint activity. This quantitative analysis reveals that neither RFC(Rad24) and the 9-1-1 clamp nor the alternative clamp loader RFC(Elg1) is required to signal paused forks. In contrast, we found that RFC(Ctf18) is essential for the Mrc1-dependent activation of Rad53 and for the maintenance of paused forks. These data identify RFC(Ctf18) as a key DRC mediator, potentially bridging Mrc1 and primed ssDNA to signal paused forks.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carrier Proteins / metabolism
Carrier Proteins / physiology
Cell Cycle Proteins / metabolism
Cell Cycle Proteins / physiology
DNA Damage
DNA Replication / drug effects
DNA Replication / physiology*
DNA, Single-Stranded / physiology
Hydroxyurea / pharmacology
Intracellular Signaling Peptides and Proteins / metabolism
Intracellular Signaling Peptides and Proteins / physiology
Saccharomyces cerevisiae / drug effects
Saccharomyces cerevisiae / genetics*
Saccharomyces cerevisiae / metabolism
Saccharomyces cerevisiae Proteins / chemistry
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism
Saccharomyces cerevisiae Proteins / physiology*
Stress, Physiological

Substances

CTF18 protein, S cerevisiae
Carrier Proteins
Cell Cycle Proteins
DNA, Single-Stranded
Elg1 protein, S cerevisiae
Intracellular Signaling Peptides and Proteins
MRC1 protein, S cerevisiae
RAD24 protein, S cerevisiae
Saccharomyces cerevisiae Proteins
rad9 protein
Hydroxyurea

Associated data

GEO/GSE21014