title: An Updated Collection of Sequence Barcoded Temperature-Sensitive Alleles of Yeast Essential Genes.
abstract: Systematic analyses of essential gene function using mutant collections in Saccharomyces cerevisiae have been conducted using collections of heterozygous diploids, promoter shut-off alleles, through alleles with destabilized mRNA, destabilized protein, or bearing mutations that lead to a temperature-sensitive (ts) phenotype. We previously described a method for construction of barcoded ts alleles in a systematic fashion. Here we report the completion of this collection of alleles covering 600 essential yeast genes. This resource covers a larger gene repertoire than previous collections and provides a complementary set of strains suitable for single gene and genomic analyses. We use deep sequencing to characterize the amino acid changes leading to the ts phenotype in half of the alleles. We also use high-throughput approaches to describe the relative ts behavior of the alleles. Finally, we demonstrate the experimental usefulness of the collection in a high-content, functional genomic screen for ts alleles that increase spontaneous P-body formation. By increasing the number of alleles and improving the annotation, this ts collection will serve as a community resource for probing new aspects of biology for essential yeast genes.
authors.affiliation: Michael Smith Laboratories, University of British Columbia, Vancouver V6T 1Z4, Canada.
authors.forename: Megan
authors.initials: M
authors.lastname: Kofoed
authors.affiliation: Terry Fox Laboratory, BC Cancer Research Centre, Vancouver, Canada.
authors.forename: Karissa L
authors.initials: KL
authors.lastname: Milbury
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver V6T 1Z4, Canada.
authors.forename: Jennifer H
authors.initials: JH
authors.lastname: Chiang
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver V6T 1Z4, Canada.
authors.forename: Sunita
authors.initials: S
authors.lastname: Sinha
authors.affiliation: Faculty of Life Sciences Bar-Ilan University, Ramat-Gan, Israel.
authors.forename: Shay
authors.initials: S
authors.lastname: Ben-Aroya
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver V6T 1Z4, Canada.
authors.forename: Guri
authors.initials: G
authors.lastname: Giaever
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver V6T 1Z4, Canada.
authors.forename: Corey
authors.initials: C
authors.lastname: Nislow
authors.affiliation: Michael Smith Laboratories, University of British Columbia, Vancouver V6T 1Z4, Canada Department of Medical Genetics, University of British Columbia, Vancouver V6T 1Z4, Canada Hieter@msl.ubc.ca pstirling@bccrc.ca.
authors.forename: Philip
authors.initials: P
authors.lastname: Hieter
authors.affiliation: Terry Fox Laboratory, BC Cancer Research Centre, Vancouver, Canada Department of Medical Genetics, University of British Columbia, Vancouver V6T 1Z4, Canada Hieter@msl.ubc.ca pstirling@bccrc.ca.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Stirling
mesh_terms: D000483:Alleles; D058893:DNA Barcoding, Taxonomic; D004171:Diploidy; D020043:Genes, Essential; D023281:Genomics; D059014:High-Throughput Nucleotide Sequencing; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins
keywords: Saccharomyces cerevisiae; essential genes; genomics; temperature-sensitive alleles