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doi:10.1534/genetics.104.035204
A more recent version of this article appeared on February 1, 2005.
REGULAR RESEARCH PAPERS |
Distinct roles for the Saccharomyces cerevisiae mismatch repair proteins in heteroduplex rejection, mismatch repair, and non-homologous tail removal
Tamara Goldfarb 1 and Eric Alani 1*
1 Cornell University
* To whom correspondence should be addressed. E-mail: eea3{at}cornell.edu.
Submitted on August 20, 2004
Revised on September 27, 2004
Accepted on 15 October 2004
The Saccharomyces cerevisiae mismatch repair (MMR) protein MSH6 and the SGS1 helicase were recently shown to play similarly important roles in preventing recombination between divergent DNA sequences in a single-strand annealing (SSA) assay. In contrast, MMR factors such as Mlh1p, Pms1p, and Exo1p were shown to not be required, or to play only minimal roles. In this study we tested mutations that disrupt Sgs1p helicase activity and Msh2p-Msh6p mismatch recognition and ATP binding and hydrolysis activities for their effect on preventing recombination between divergent DNA sequences (heteroduplex rejection) during SSA. The results support a model in which the Msh proteins act with Sgs1p to unwind DNA recombination intermediates containing mismatches. Importantly, msh2 mutants were characterized that displayed separation of function phenotypes with respect to non-homologous tail removal during SSA and heteroduplex rejection. These studies suggest that non-homologous tail removal is a separate function of Msh proteins that is likely to involve a distinct DNA binding activity. The involvement of Sgs1p in heteroduplex rejection but not non-homologous tail removal further illustrates that subsets of MMR proteins collaborate with factors in different DNA repair pathways to maintain genome stability.
Key Words: heteroduplex rejection, mismatch repair, non-homologous tail removal
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