A genome-wide view of mutations in respiration-deficient mutants of Saccharomyces cerevisiae selected following carbon ion beam irradiation
Mitochondrial dysfunction in Saccharomyces cerevisiae was selected as a marker of ion penetration following carbon ion beam (CIB) irradiation. Respiration-deficient mutants were screened. Following confirmation of negligible spontaneous mutation, eight genetically stable S. cerevisiae respiration-deficient mutant strains and a control strain were resequenced with ~ 200-fold read depth. Strategies were established to identify and validate the particular mutations induced by CIB irradiation. In the nuclear genome, CIB irradiation mainly caused base substitutions and some small (< 100 bp) insertions/deletions (indels), which were widely distributed across the chromosomes. Although mitochondrial dysfunction was selected as a screening marker, variants in the nuclear genome were detected at a high frequency (10−7) relative to spontaneous mutations (10−9). The transition to transversion ratio for base substitutions was 0.746, which was less than that of spontaneous mutations. In the mitochondrial genome, there were very large deletions including substantial gene areas, resulting in extremely low read coverage. Meanwhile, every mutant possessed a distinctive mutation pattern, for both the nuclear and the mitochondrial genome. Nuclear genomes contained scanty mitochondrial respiration-related genes that were potentially affected by verified mutations, suggesting that variants in the mitochondrial genome may be the main drivers of respiratory deficiencies. Our study confirmed the previous finding that heavy ion beam (HIB) irradiation mainly induces substantial base substitutions and some small indels but also yielded some novel findings, in particular, novel structural variants in the mitochondrial genomes. These data will enhance the understanding of HIB-induced damage and mutations and aid in the HIB-based microbial mutation breeding.
KeywordsSaccharomyces cerevisiae Carbon ion beam irradiation Respiration-deficient mutants Genome-wide resequencing Molecular mutation spectrum Mitochondrial genome
The authors would like to thank the colleagues at HIRFL for providing high-quality carbon ion beam irradiation.
This work was funded by the Chinese Academy of Sciences Key Deployment Project (No. KFZD-SW-109), a Joint project of the Chinese Academy of Sciences and the Industrial Technology Research Institute (CAS-ITRI 201801) and the National Natural Science Fund of China (No. 11575259).
Compliance with ethical standards
Conflict of interest
The authors declare that there are no conflicts of interest.
Human and animal rights
This article does not contain any studies with human participants or animals performed by any of the authors.
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