SUMMARY We report the construction and analysis of 4,836 heterozygous diploid deletion mutants covering 98.4% of the fission yeast genome. This resource provides a powerful tool for biotechnological and eukaryotic cell biology research. Comprehensive gene dispensability comparisons with budding yeast, the first time such studies have been possible between two eukaryotes, revealed that 83% of single copy orthologues in the two yeasts had conserved dispensability. Gene dispensability differed for certain pathways between the two yeasts, including mitochondrial translation and cell cycle checkpoint control. We show that fission yeast has more essential genes than budding yeast and that essential genes are more likely than non-essential genes to be single copy, broadly conserved and to contain introns. Growth fitness analyses determined sets of haploinsufficient and haploproficient genes for fission yeast, and comparisons with budding yeast identified specific ribosomal proteins and RNA polymerase subunits, which may act more generally to regulate eukaryotic cell growth.

An epistasis map (E-MAP) was constructed in the fission yeast, Schizosaccharomyces pombe, by systematically measuring the phenotypes associated with pairs of mutations. This high-density, quantitative genetic interaction map focused on various aspects of chromosome function, including transcription regulation and DNA repair/replication. The E-MAP uncovered a previously unidentified component of the RNA interference (RNAi) machinery (rsh1) and linked the RNAi pathway to several other biological processes. Comparison of the S. pombe E-MAP to an analogous genetic map from the budding yeast revealed that, whereas negative interactions were conserved between genes involved in similar biological processes, positive interactions and overall genetic profiles between pairs of genes coding for physically associated proteins were even more conserved. Hence, conservation occurs at the level of the functional module (protein complex), but the genetic cross talk between modules can differ substantially.Genetic interactions report on the extent to which the function of one gene depends on the presence of a second. This phenomenon, known as epistasis, can be used for defining functional relationships between genes and the pathways in which the corresponding proteins function. Two main categories of genetic interactions exist: (i) negative (e.g., synthetic sickness/ lethality) and (ii) positive (e.g., suppression). We have developed a quantitative approach, termed epistasis map (E-MAP), allowing us to measure the whole spectrum of genetic interactions, both positive and negative (1,2). In budding yeast, Saccharomyces cerevisiae, it has been demonstrated that positive genetic interactions can identify pairs of genes whose products are physically associated and/or function in the same pathway (1,2), whereas negative interactions exist between genes acting on parallel pathways (3,4).We developed the Pombe Epistasis Mapper (PEM) approach (5) that allows high-throughput generation of double mutants in the fission yeast, Schizosaccharomyces pombe. Fission yeast is more similar to metazoans than is S. cerevisiae, owing to its large complex centromere structure, the restriction of spindle construction to mitotic entry, gene regulation by histone methylation and chromodomain heterochromatin proteins, gene and transposon regulation by the RNA interference (RNAi) pathway, and the widespread presence of introns in genes. To further study these processes and to try to understand how genetic interaction networks have evolved (6), we generated an E-MAP in S. pombe that focuses on nuclear function, designed to be analogous to one we created in budding yeast (2). An E-MAP in S. pombeUsing our PEM system (5), we generated a quantitative genetic interaction map in S. pombe, comprising ~118,000 distinct double mutant combinations among 550 genes involved in various aspects of chromosome function ( Fig. 1A and tables S1 and S4) (7). The genes on the map were chosen on the basis of a previous budding yeast E-MAP (1,2) and also included factor...