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PMID: 11095674 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Alteration of gene conversion tract length and associated crossing over during plasmid gap repair in nuclease-deficient strains of Saccharomyces cerevisiae.

Nucleic acids research ·Vol. 28 ·No. 23 ·2000-12-01 ·Pages 4649-56

Symington LS, Kang LE, Moreau S

Abstract

A plasmid gap repair assay was used to assess the role of three known nucleases, Exo1, Mre11 and Rad1, in the processing of DNA ends and resolution of recombination intermediates during double-strand gap repair. In this assay, alterations in end processing or branch migration are reflected by the frequency of co-conversion of a chromosomal marker 200 bp from the gap. Gap repair associated with crossing over results in integration at the homologous chromosomal locus, whereas the plasmid remains episomal for non-crossover repair events. In mre11 strains, the frequency of gap repair was reduced 3- to 10-fold and conversion tracts were shorter than in the wild-type strain, consistent with a role for this nuclease in processing double-strand breaks. However, conversion tracts were longer in a strain containing the nuclease deficient allele, mre11-H125N, suggesting increased end processing by redundant nucleases. The frequency of gap repair was reduced 2-fold in rad1 mutants and crossing over was reduced, consistent with a role for Rad1 in cleaving recombination intermediates. The frequency of gap repair was increased in exo1 mutants with a significant increase in crossing over. In exo1 mre11 double mutants gap repair was reduced to below the mre11 single mutant level.

MeSH Terms
Crossing Over, Genetic/genetics DNA Repair DNA Repair Enzymes DNA-Binding Proteins Deoxyribonucleases/deficiency,genetics Endodeoxyribonucleases Endonucleases/deficiency,genetics Exodeoxyribonucleases/deficiency,genetics Fungal Proteins/genetics,metabolism Gene Conversion/genetics Mutation Plasmids/genetics,metabolism Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins
Chemicals
DNA-Binding Proteins Fungal Proteins Saccharomyces cerevisiae Proteins Deoxyribonucleases Endodeoxyribonucleases Endonucleases Exodeoxyribonucleases MRE11 protein, S cerevisiae RAD1 protein, S cerevisiae exodeoxyribonuclease I DNA Repair Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Symington L S
Department of Microbiology and Institute of Cancer Research, Columbia University College of Physicians and Surgeons, 701 West 168th Street, New York, NY 10032, USA. lss5@columbia.edu
Kang L E
Moreau S
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/28.23.4649
Email
lss5@columbia.edu
Published
2000-12-01
Pages
4649-56
Language
English
Region
England
NLM ID
0411011
PMCID
PMC115160
Subset
IM
Grants
NIGMS NIH HHS · R01 GM041784 · United States
NIGMS NIH HHS · R01 GM054099 · United States
NIGMS NIH HHS · GM54099 · United States
External Links
PubMed source DOI: 10.1093/nar/28.23.4649 PMC Full Text View journal details
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