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PMID: 12912929 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

DNA polymerase stabilization at stalled replication forks requires Mec1 and the RecQ helicase Sgs1.

The EMBO journal ·Vol. 22 ·No. 16 ·2003-08-15 ·Pages 4325-36

Cobb JA, Bjergbaek L, Shimada K, Frei C, Gasser SM

Abstract

To ensure proper replication and segregation of the genome, eukaryotic cells have evolved surveillance systems that monitor and react to impaired replication fork progression. In budding yeast, the intra-S phase checkpoint responds to stalled replication forks by downregulating late-firing origins, preventing spindle elongation and allowing efficient resumption of DNA synthesis after recovery from stress. Mutations in this pathway lead to high levels of genomic instability, particularly in the presence of DNA damage. Here we demonstrate by chromatin immunoprecipitation that when yeast replication forks stall due to hydroxyurea (HU) treatment, DNA polymerases alpha and epsilon are stabilized for 40-60 min. This requires the activities of Sgs1, a member of the RecQ family of DNA helicases, and the ATM-related kinase Mec1, but not Rad53 activation. A model is proposed whereby Sgs1 helicase resolves aberrantly paired structures at stalled forks to maintain single-stranded DNA that allows RP-A and Mec1 to promote DNA polymerase association.

MeSH Terms
DNA Helicases/chemistry,genetics,metabolism DNA Repair DNA, Fungal/drug effects DNA-Directed DNA Polymerase/metabolism Hydroxyurea/pharmacology Intracellular Signaling Peptides and Proteins Kinetics Models, Biological Nucleic Acid Synthesis Inhibitors/pharmacology Point Mutation Protein Serine-Threonine Kinases Protein Structure, Tertiary RecQ Helicases Replication Origin S Phase Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/metabolism Time Factors
Chemicals
DNA, Fungal Intracellular Signaling Peptides and Proteins Nucleic Acid Synthesis Inhibitors Saccharomyces cerevisiae Proteins MEC1 protein, S cerevisiae Protein Serine-Threonine Kinases DNA-Directed DNA Polymerase SGS1 protein, S cerevisiae DNA Helicases RecQ Helicases Hydroxyurea
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Cobb Jennifer A
University of Geneva, Department of Molecular Biology, Quai Ernest-Ansermet 30, CH-1211 Geneva, Switzerland.
Bjergbaek Lotte
Shimada Kenji
Frei Christian
Gasser Susan M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg391
Published
2003-08-15
Pages
4325-36
Language
English
Region
England
NLM ID
8208664
PMCID
PMC175781
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/cdg391 PMC Full Text View journal details
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