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Synthetic hypermutation : gene-drug mutation rate synergy reveals a translesion synthesis mechanism

University of British Columbia

Gene-gene or gene-drug interactions are typically quantified using fitness as a readout because the data is continuous and easily measured in high-throughput. However, to what extent fitness captures the range of other phenotypes that show synergistic effects is usually unknown. Using Saccharomyces cerevisiae, and focusing on a matrix of DNA repair mutants and genotoxic drugs, I quantified 76 gene-drug interactions based on both mutation rate and fitness and find that these parameters are not connected. Independent of fitness defects I identified seven cases of synthetic hypermutation, where the combined effect of the drug and mutant on mutation rate was greater than predicted. One example occurred when yeast lacking RAD1 were exposed to cisplatin and I characterized this interaction using whole-genome sequencing. Our sequencing results indicate mutagenesis by cisplatin in rad1∆ cells depended almost entirely on interstrand crosslinks at GpCpN motifs. Interestingly, our data suggest that the 3’ base in this motif templates the addition of the mutated base. This result differs from cisplatin mutation signatures in XPF-deficient C. elegans and supports a model in which translesion synthesis polymerases perform a slippage and realignment extension across from the damaged base. Accordingly, DNA polymerase zeta activity was essential for mutagenesis in cisplatin-treated rad1∆ cells. Together these data reveal the potential to gain new mechanistic insights from non-fitness measures of gene-drug interactions and extend the use of mutation accumulation and whole-genome sequencing analysis to define DNA repair mechanisms.

Text

2017-04-19

Attribution-NonCommercial-NoDerivatives 4.0 International

http://hdl.handle.net/2429/61269

Medical Genetics

University of British Columbia

UBCV

http://creativecommons.org/licenses/by-nc-nd/4.0/