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PMID: 21980306 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Genome-wide analysis of heteroduplex DNA in mismatch repair-deficient yeast cells reveals novel properties of meiotic recombination pathways.

PLoS genetics ·Vol. 7 ·No. 9 ·2011-09-00 ·Pages e1002305

Martini E, Borde V, Legendre M, Audic S, Regnault B, Soubigou G, Dujon B, Llorente B

Abstract

Meiotic DNA double-strand breaks (DSBs) initiate crossover (CO) recombination, which is necessary for accurate chromosome segregation, but DSBs may also repair as non-crossovers (NCOs). Multiple recombination pathways with specific intermediates are expected to lead to COs and NCOs. We revisited the mechanisms of meiotic DSB repair and the regulation of CO formation, by conducting a genome-wide analysis of strand-transfer intermediates associated with recombination events. We performed this analysis in a SK1 × S288C Saccharomyces cerevisiae hybrid lacking the mismatch repair (MMR) protein Msh2, to allow efficient detection of heteroduplex DNAs (hDNAs). First, we observed that the anti-recombinogenic activity of MMR is responsible for a 20% drop in CO number, suggesting that in MMR-proficient cells some DSBs are repaired using the sister chromatid as a template when polymorphisms are present. Second, we observed that a large fraction of NCOs were associated with trans-hDNA tracts constrained to a single chromatid. This unexpected finding is compatible with dissolution of double Holliday junctions (dHJs) during repair, and it suggests the existence of a novel control point for CO formation at the level of the dHJ intermediate, in addition to the previously described control point before the dHJ formation step. Finally, we observed that COs are associated with complex hDNA patterns, confirming that the canonical double-strand break repair model is not sufficient to explain the formation of most COs. We propose that multiple factors contribute to the complexity of recombination intermediates. These factors include repair of nicks and double-stranded gaps, template switches between non-sister and sister chromatids, and HJ branch migration. Finally, the good correlation between the strand transfer properties observed in the absence of and in the presence of Msh2 suggests that the intermediates detected in the absence of Msh2 reflect normal intermediates.

MeSH Terms
Chromatids/genetics Chromosome Segregation Crossing Over, Genetic DNA Breaks, Double-Stranded DNA Mismatch Repair/genetics DNA Repair/genetics DNA, Cruciform/genetics Genome-Wide Association Study Meiosis/genetics MutS Homolog 2 Protein/genetics Nucleic Acid Heteroduplexes/genetics Recombination, Genetic Saccharomyces cerevisiae/cytology,genetics,metabolism Saccharomyces cerevisiae Proteins/genetics Sister Chromatid Exchange
Chemicals
DNA, Cruciform Nucleic Acid Heteroduplexes Saccharomyces cerevisiae Proteins MSH2 protein, S cerevisiae MutS Homolog 2 Protein
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Martini Emmanuelle
CEA DSV/IRCM, Unité Mixte de Recherche 217 Radiobiologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Commissariat à l'Energie Atomique et aux Energies Alternatives, Fontenay aux Roses, France. [email protected]
Borde Valérie
Legendre Matthieu
Audic Stéphane
Regnault Béatrice
Soubigou Guillaume
Dujon Bernard
Llorente Bertrand
Conflict of Interest

The authors have declared that no competing interests exist.

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Article Info
Journal
PLoS genetics
Abbr.
PLoS Genet
ISSN
1553-7404
DOI
10.1371/journal.pgen.1002305
Email
[email protected]
Published
2011-09-00
Epub
2011-00-29
Pages
e1002305
Language
English
Region
United States
NLM ID
101239074
PMCID
PMC3183076
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pgen.1002305 PMC Full Text View journal details
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