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Suppression of homeologous recombination by mismatch repair and nucleotide excision repair proteins in Saccharomyces cerevisiae
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Suppression of homeologous recombination by mismatch repair and nucleotide excision repair proteins in Saccharomyces cerevisiae
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Suppression of homeologous recombination by mismatch repair and nucleotide excision repair proteins in Saccharomyces cerevisiae
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Dissertation
Suppression of homeologous recombination by mismatch repair and nucleotide excision repair proteins in Saccharomyces cerevisiae
2000
Overview
Mismatch repair proteins suppress recombination between sequences that are similar but non-identical (homeologous). In the yeast Saccharomyces cerevisiae, the nucleotide excision repair proteins Rad1p and Rad10p interact with mismatch repair proteins Msh2p and Msh3p at certain stages of recombination, including processing of terminal non-homology when necessary. Two recombination assays were used to examine the roles of DNA repair proteins in suppression of homeologous recombination and to study the interactions between mismatch repair and nucleotide excision repair proteins. A previously developed inverted repeat assay was modified and used to confirm that the specificities of mismatch recognition of the MutS homologs in recombination are similar to their specificities in DNA repair, with the surprising exception that Msh3p appears to recognize base-base mismatches during recombination. Roles of MutS homologs in recombination suppression independent of MutL homologs were indicated, and the Rad1p/Rad10p heterodimer was found to act in suppression of homeologous recombination. A novel ectopic recombination assay was developed that can place recombination substrates in competition, allowing measurement of the strength of preference for homologous over homeologous substrates. This assay was also used to examine the effect of terminal non-homology on recombination rates. We found that the presence of even small amounts of terminal non-homology inhibits recombination despite the presence of all proteins necessary for its processing, and that disruption of MSH2, MSH3, or RAD1 further reduces the ability of cells to recombine when terminal non-homology must be processed. All recognition of base-base mismatches in this assay proceeded via a Msh2p/Msh6p heterodimer, and 4nt loops did not inhibit recombination.
Publisher
ProQuest Dissertations & Theses
Subject
ISBN
9780599889965, 0599889969
