Abstract
Synthetic genetic array (SGA) has been successfully used to identify genetic interactions in S. cerevisiae and S. pombe. In S. pombe, SGA methods use either cycloheximide (C) or heat shock (HS) to select double mutants before measuring colony size as a surrogate for fitness. Quantitative Fitness Analysis (QFA) is a different method for determining fitness of microbial strains. In QFA, liquid cultures are spotted onto solid agar and growth curves determined for each spot by photography and model fitting. Here, we compared the two S. pombe SGA methods and found that the HS method was more reproducible for us. We also developed a QFA procedure for S. pombe. We used QFA to identify genetic interactions affecting two temperature sensitive, telomere associated query mutations (taz1Δ and pot1-1). We identify exo1∆ and other gene deletions as suppressors or enhancers of S. pombe telomere defects. Our study identifies known and novel gene deletions affecting the fitness of strains with telomere defects. The interactions we identify may be relevant in human cells.
Highlights
Genetic interactions (GIs) arise when the function of one gene is affected by the function of another [1]
Each of the two S. pombe synthetic genetic array (SGA) methods to generate double mutants should result in a similar pattern of GIs
We found that the fitness rankings for his3Δ, ura5Δ and his7Δ SGAs were different to each other in the rich media we use for SGA
Summary
Genetic interactions (GIs) arise when the function of one gene is affected by the function of another [1]. Synthetic genetic array (SGA) methodology has been used to characterise GIs on a genome-wide scale [2,3,4]. SGA uses large-scale robotic procedures for mating and sporulation carried out on solid agar media to generate double mutant colonies and to measure their size [5]. By comparing the size of double mutants it is possible to classify GIs as negative (where double mutant colonies are smaller than expected), positive (larger than expected) or neutral [6,7,8,9,10]. Genome-wide SGA screens performed using the budding yeast S.
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