Mismatch Correction Acts as a Barrier to Homeologous Recombination in Saccharomyces cerevisiae

INVESTIGATIONS

Mismatch Correction Acts as a Barrier to Homeologous Recombination in Saccharomyces cerevisiae

E. M. Selva, L. New, G. F. Crouse and R. S. Lahue
Department of Biochemistry and Molecular Biology, University of Massachusetts Medical Center, Worcester, Massachusetts 01655

A homeologous mitotic recombination assay was used to test the role of Saccharomyces cerevisiae mismatch repair genes PMS1, MSH2 and MSH3 on recombination fidelity. A homeologous gene pair consisting of S. cerevisiae SPT15 and its S. pombe homolog were present as a direct repeat on chromosome V, with the exogenous S. pombe sequences inserted either upstream or downstream of the endogenous S. cerevisiae gene. Each gene carried a different inactivating mutation, rendering the starting strain Spt15(-). Recombinants that regenerated SPT15 function were scored after nonselective growth of the cells. In strains wild type for mismatch repair, homeologous recombination was depressed 150- to 180-fold relative to homologous controls, indicating that recombination between diverged sequences is inhibited. In one orientation of the homeologous gene pair, msh2 or msh3 mutations resulted in 17- and 9.6-fold elevations in recombination and the msh2 msh3 double mutant exhibited an 43-fold increase, implying that each MSH gene can function independently in trans to prevent homeologous recombination. Homologous recombination was not significantly affected by the msh mutations. In the other orientation, only msh2 strains were elevated (12-fold) for homeologous recombination. A mutation in MSH3 did not affect the rate of recombination in this orientation. Surprisingly, a pms1 deletion mutant did not exhibit elevated homeologous recombination.

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				N. Sugawara, T. Goldfarb, B. Studamire, E. Alani, and J. E. Haber Heteroduplex rejection during single-strand annealing requires Sgs1 helicase and mismatch repair proteins Msh2 and Msh6 but not Pms1 PNAS, June 22, 2004; 101(25): 9315 - 9320. [Abstract] [Full Text] [PDF]

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				B. Studamire, G. Price, N. Sugawara, J. E. Haber, and E. Alani Separation-of-Function Mutations in Saccharomyces cerevisiae MSH2 That Confer Mismatch Repair Defects but Do Not Affect Nonhomologous-Tail Removal during Recombination Mol. Cell. Biol., November 1, 1999; 19(11): 7558 - 7567. [Abstract] [Full Text] [PDF]

				J. A. Nickoloff, D. B. Sweetser, J. A. Clikeman, G. J. Khalsa, and S. L. Wheeler Multiple Heterologies Increase Mitotic Double-Strand Break-Induced Allelic Gene Conversion Tract Lengths in Yeast Genetics, October 1, 1999; 153(2): 665 - 679. [Abstract] [Full Text] [PDF]

				C. E. Schrader, W. Edelmann, R. Kucherlapati, and J. Stavnezer Reduced Isotype Switching in Splenic B Cells from Mice Deficient in Mismatch Repair Enzymes J. Exp. Med., August 2, 1999; 190(3): 323 - 330. [Abstract] [Full Text] [PDF]

				O. Inbar and M. Kupiec Homology Search and Choice of Homologous Partner during Mitotic Recombination Mol. Cell. Biol., June 1, 1999; 19(6): 4134 - 4142. [Abstract] [Full Text] [PDF]

				F. Paques and J. E. Haber Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae Microbiol. Mol. Biol. Rev., June 1, 1999; 63(2): 349 - 404. [Abstract] [Full Text] [PDF]

				W. Chen and S. Jinks-Robertson The Role of the Mismatch Repair Machinery in Regulating Mitotic and Meiotic Recombination Between Diverged Sequences in Yeast Genetics, April 1, 1999; 151(4): 1299 - 1313. [Abstract] [Full Text]