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Originally published as Genetics Published Articles Ahead of Print on April 2, 2006.
Genetics, Vol. 173, 647-659, June 2006, Copyright © 2006
doi:10.1534/genetics.105.055244
The Effects of Mismatch Repair and RAD1 Genes on Interchromosomal Crossover Recombination in Saccharomyces cerevisiae
Ainsley Nicholson*,1,
Rebecca M. Fabbri
,
Jason W. Reeves
and
Gray F. Crouse*,
,2
* Graduate Program in Genetics and Molecular Biology and
Department of Biology, Emory University, Atlanta, Georgia 30322
2 Corresponding author: Department of Biology, 1510 Clifton Rd., Atlanta, GA 30322.
E-mail: gcrouse{at}biology.emory.edu
We have previously shown that recombination between 400-bp substrates containing only 4-bp differences, when present in an inverted repeat orientation, is suppressed by >20-fold in wild-type strains of S. cerevisiae. Among the genes involved in this suppression were three genes involved in mismatch repairMSH2, MSH3, and MSH6and one in nucleotide excision repair, RAD1. We now report the involvement of these genes in interchromosomal recombination occurring via crossovers using these same short substrates. In these experiments, recombination was stimulated by a double-strand break generated by the HO endonuclease and can occur between completely identical (homologous) substrates or between nonidentical (homeologous) substrates. In addition, a unique feature of this system is that recombining DNA strands can be given a choice of either type of substrate. We find that interchromosomal crossover recombination with these short substrates is severely inhibited in the absence of MSH2, MSH3, or RAD1 and is relatively insensitive to the presence of mismatches. We propose that crossover recombination with these short substrates requires the products of MSH2, MSH3, and RAD1 and that these proteins have functions in recombination in addition to the removal of terminal nonhomology. We further propose that the observed insensitivity to homeology is a result of the difference in recombinational mechanism and/or the timing of the observed recombination events. These results are in contrast with those obtained using longer substrates and may be particularly relevant to recombination events between the abundant short repeated sequences that characterize the genomes of higher eukaryotes.
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