The DNA damage checkpoint pathways exert multiple controls on the efficiency and outcome of the repair of a double-stranded DNA gap

Haghnazari, E.

doi:10.1093/nar/gkh717

Cited by 20DNA Repair MechanismsFungal and yeast genetics researchBacterial Genetics and Biotechnology

The DNA damage checkpoint pathways exert multiple controls on the efficiency and outcome of the repair of a double-stranded DNA gap

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Haghnazari, E.·2004·Nucleic Acids Research

DOI 10.1093/nar/gkh717PMC PMC514360

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Abstract

A DNA gap repair assay was used to determine the effect of mutations in the DNA damage checkpoint system on the efficiency and outcome (crossover/non-crossover) of recombinational DNA repair. In Saccharomyces cerevisiae gap repair is largely achieved by homologous recombination. As a result the plasmid either integrates into the chromosome (indicative of a crossover outcome) or remains extrachromosomal (indicative of a non-crossover outcome). Deletion mutants of the MEC1 and RAD53 checkpoint kinase genes exhibited a 5-fold decrease in gap repair efficiency, showing that 80% of the gap repair events depended on functional DNA damage checkpoints. Epistasis analysis suggests that the DNA damage checkpoints affect gap repair by modulating Rad51 protein-mediated homologous recombination. While in wild-type cells only approximately 25% of the gap repair events were associated with a crossover outcome, Mec1-deficient cells exhibited a >80% crossover association. Also mutations in the effector kinases Rad53, Chk1 and Dun1 were found to affect crossover association of DNA gap repair to various degrees. The data suggest that the DNA damage checkpoints are important for the optimal functioning of recombinational DNA repair with multiple terminal targets to modulate the efficiency and outcome of homologous recombination.

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In yeast, deletion of MEC1 and Rad53 checkpoint kinases or downstream effector kinases dramatically decreases double-strand break repair efficiency.
“Similarly, such a checkpoint-dependent regulation of double-strand break repair has been observed in yeast where deletion of the MEC1 and Rad53 checkpoint kinases or downstream effector kinases led to a dramatic decrease in double-strand break repair efficiency (29,30).”
From Phosphorylation of Nucleotide Excision Repair Factor Xeroderma Pigmentosum Group A by Ataxia Telangiectasia Mutated and Rad3-Related–Dependent Checkpoint Pathway Promotes Cell Survival in Response to UV Irradiation (2006) · cites this paper as [30]
In “Mec1 and Rad53 kinases are required for efficient gap repair” (page not provided), the paper reports a fivefold reduction in double-strand gap repair in mec1 and rad53 mutants. But it also reports that the downstream kinase mutants dun1 and chk1 did not differ significantly from wild type in repair frequency, so the claim overextends that finding to downstream effector kinases. (LLM, conf=0.90)
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Authors: Haghnazari, E.
Affiliations: University of California, Davis (US)
Corresponding author: Haghnazari, E.
Venue: Nucleic Acids Research
Publisher: Oxford University Press (OUP)
Published: 2004-08-03
Volume: 32
Issue: 14
Pages: 4257-4268
ISSN: 0305-1048 (ISSN-L)
1362-4962
DOI: 10.1093/nar/gkh717
PMC: PMC514360
OpenAlex: W2125132000
Cited by: 20
References: 91
Citations by year: 2020: 1 · 2016: 1 · 2015: 1 · 2014: 1 · 2013: 2
FWCI: 1.07
Type: journal article
Language: en
Open access: bronze
doi.org
Locations: Nucleic Acids Research (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health
National Cancer Institute (CA92276, R01CA092276)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords: DNA damage checkpoint, homologous recombination, Mec1, Rad53, Chk1, Dun1, RAD51, epistasis analysis, plasmid integration, Saccharomyces cerevisiae
MeSH terms: DNA Damage, DNA Repair, DNA-Binding Proteins, Mitosis, Mutation, Recombination, Genetic, Saccharomyces cerevisiae, Transformation, Genetic, Signal Transduction, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Saccharomyces cerevisiae Proteins, Intracellular Signaling Peptides and Proteins, Rad51 Recombinase, Checkpoint Kinase 2

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The DNA damage checkpoint pathways exert multiple controls on the efficiency and outcome of the repair of a double-stranded DNA gap

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