Expression of Saccharomyces cerevisiae MATa and MAT alpha enhances the HO endonuclease-stimulation of chromosomal rearrangements directed by his3 recombinational substrates.
Radiation resistance in Saccharomyces cerevisiae is greater in a/alpha diploids than in aa or alpha alpha diploids, and higher levels of radiation resistance correlates with more mitotic recombination. Specifically, we investigated whether the stimulation of directed translocations, inversions, and unequal sister chromatid exchanges (SCEs) by HO endonuclease-induced double-strand breaks (DSBs) is enhanced in a/alpha cells. These rearrangements result from mitotic recombination between two truncated his3 genes, his3-delta 5' and his3-delta 3'::HOcs, positioned on non-homologous chromosomes or positioned in juxtaposition on the same chromosome in inverted or direct orientation. Mitotic recombination was initiated by HO endonuclease-induced DSBs at the HO cut site (HOcs) located at his3-delta 3'::HOcs, and His+ recombinants were selected. In MATa-inc haploid strains, which do not switch mating-type, the DSB reduced viability, relative to undamaged cells, and increases the frequency of His+ recombinants containing translocations to 2.4 x 10(-4) (seven-fold), SCEs to 5.4 x 10(-4) (five-fold), and inversions to 1.8 x 10(-3) (six-fold). Compared to a haploids, DSB-stimulated frequencies in a/alpha haploids were three-fold higher for translocations, two-fold higher for SCEs, and ten-fold higher for inversions; however DSB-induced lethality was greater in a/alpha haploids. Compared to aa diploids, DSB-stimulated frequencies of translocations and viability after chromosome cleavage were greater in a/alpha diploids. We suggest that heterozygosity at MAT may elevate the frequency of DSB-initiated reciprocal exchange events in both haploid and diploid cells, but may only increase viability after chromosome cleavage in diploid cells.
Fasullo
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Department of Biochemistry and Molecular Biology, Albany Medical College, NY 12208, USA. mfasullo@ccgateway.amc.edu
Bennett
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Dave
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D019457:Chromosome Breakage / Q000235:genetics; D002875:Chromosomes / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D049770:Genes, Mating Type, Fungal; D006836:Hydro-Lyases / Q000235:genetics*; D008938:Mitosis / Q000235:genetics; D011003:Ploidies; D011995:Recombination, Genetic / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D012854:Sister Chromatid Exchange / Q000235:genetics; D014178:Translocation, Genetic / Q000235:genetics
D029701:Saccharomyces cerevisiae Proteins; C530845:HO protein, S cerevisiae; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase