Abstract
Mutations in budding yeast occur in meiosis at higher frequencies than in cells grown vegetatively. In contrast to mutations that occur in somatic cells, meiotic mutations have a special, long-range impact on evolution, because they are transferred to the following generations through the gametes. Understanding the mechanistic basis of meiotic mutagenicity is still lacking, however. Here, we report studies of mutagenicity in the reporter gene CAN1, in which forward mutation events in meiosis are sevenfold higher than in mitotic cells, as determined by fluctuation analysis. Meiotic mutations appear approximately at the same time as heteroallelic-recombination products and as meiotic DSBs. Recombination-associated timing of meiotic mutagenicity is further augmented by the absence of meiotic mutations in cells arrested after pre-meiotic DNA synthesis. More than 40% of the mutations generated in meiosis in CAN1 are found on chromosomes that have recombined in the 2.2 kb covering the reporter, implying that the mutations have resulted from recombination events and that meiotic recombination is mutagenic. The induced expression in yeast meiosis of low-fidelity DNA polymerases coded by the genes REV1, REV3, RAD30, and POL4 makes them attractive candidates for introducing mutations. However, in our extensive experiments with polymerase-deleted strains, these polymerases do not appear to be the major source of meiotic mutagenicity. From the connection between meiotic mutagenicity and recombination, one may conclude that meiotic recombination has another diversification role, of introducing new mutations at the DNA sequence level, in addition to reshuffling of existing variation. The new, rare meiotic mutations may contribute to long-range evolutionary processes and enhance adaptation to challenging environments.
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Acknowledgements
We thank Dr. Nancy Hollingsworth for providing the cdc7-as3 strains, and Dr. Alex Levitzki and Dr. Shoshana Klein for supplying the PP1 inhibitor. Our work was supported by funds provided by the Israel Science Foundation (Grant # 501/16), the US–Israel Binational Science Foundation (Grant # 2009299), and by Hadassah Academic College.
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Conceived and designed the experiments: OM, GS, and AAE. Performed the experiments: OM, LM, KZ, and RA. Contributed reagents/materials/analysis tools: DZ. Wrote the paper: OM, GS, and AAE.
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Fig. S1
Mutations arising during vegetative growth in WT and TLSP-deleted strains following UV treatment. Exponentially growing haploid cells (a) and diploid cells (c) were serially diluted (1:10) and each dilution was spotted onto 2 YEPD plates, and incubated at 30 °C over-night. Cells from one plate with haploid cultures (a) and from one plate with diploid cultures (c) were replica plated onto Can, -Arg plates, UV irradiated (100J/m2), and incubated in the dark for 4 days at 30°C (b and d respectively) (TIF 259 kb)
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Mansour, O., Morciano, L., Zion, K. et al. Timing of appearance of new mutations during yeast meiosis and their association with recombination. Curr Genet 66, 577–592 (2020). https://doi.org/10.1007/s00294-019-01051-0
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DOI: https://doi.org/10.1007/s00294-019-01051-0