Analysis of a genome-wide set of gene deletions in the fission yeast Schizosaccharomyces pombe

Abstract

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.

The SAGA Histone Acetyltransferase Complex Regulates Leucine Uptake through the Agp3 Permease in Fission Yeast

“…The pH of each 2.2ϫ EMM2 medium with or without a nitrogen source was adjusted to 5.5. The commercially available haploid gene deletion strains (19) were purchased from Bioneer Corp. Other strains are constructed using standard techniques as described previously (20). All strains are listed in supplemental Table 1.…”

“…2), leu1 Ϫ gcn5⌬ double mutants should grow better than leu1 Ϫ mutants in the presence of excess NH 4 Cl. This idea could be tested using a haploid commercially available gene deletion library constructed in the leu1-32 ura4-D18 ade6 background (19). Therefore, we examined the growth of these mutants on minimal media containing excess NH 4 Cl and the necessary supplements.…”

Break-induced ATR and Ddb1–Cul4Cdt2 ubiquitin ligase-dependent nucleotide synthesis promotes homologous recombination repair in fission yeast

“…Following removal of thiamine (T) from the media, the HO endonuclease is expressed, inducing a unique sitespecific DSB at the MATa recognition site located on the right arm. This newly created minichromosome was introduced into a subset of 205 mutants from the S. pombe Bioneer haploid deletion library (Kim et al 2010) exhibiting sensitivity to the alkylating agent methylmethane sulfonate (MMS) and/or the radiomimetic bleomycin (Deshpande et al 2009;our unpublished results). Ch 16 encodes an ade6-M216 point mutation that, when present with an ade6-M210 heteroallele on ChIII, results in an ade + (white) phenotype through intragenic complementation (Leupold and Gutz 1964).…”

A functional genome-wide genetic screening identifies new pathways controlling the G1/S transcriptional wave

“…17 High throughput crossing of the reporter strain with the deletion library in 96-well plates followed by direct plating on selective media allowed the isolation of haploid strains, each one with a single gene deletion and the Cdc22-YFP and the psty1-RFP reporters. Of the total collection, we were able to cross and measure a YFP/FSC ratio in 2792 strains, which represents 92.9% of the original strain collection.…”