title: Functional Analysis of Kinases and Transcription Factors inSaccharomyces cerevisiaeUsing an Integrated Overexpression Library.
abstract: Kinases and transcription factors (TFs) are key modulators of important signaling pathways and their activities underlie the proper function of many basic cellular processes such as cell division, differentiation, and development. Changes in kinase and TF dosage are often associated with disease, yet a systematic assessment of the cellular phenotypes caused by the combined perturbation of kinases and TFs has not been undertaken. We used a reverse-genetics approach to study the phenotypic consequences of kinase and TF overexpression (OE) in the budding yeast,Saccharomyces cerevisiaeWe constructed a collection of strains expressing stably integrated inducible alleles of kinases and TFs and used a variety of assays to characterize the phenotypes caused by TF and kinase OE. We used the Synthetic Genetic Array (SGA) method to examine dosage-dependent genetic interactions (GIs) between 239 gain-of-function (OE) alleles of TFs and six loss-of-function (LOF) and seven OE kinase alleles, the former identifying Synthetic Dosage Lethal (SDL) interactions and the latter testing a GI we call Double Dosage Lethality (DDL). We identified and confirmed 94 GIs between 65 OE alleles of TFs and 9 kinase alleles. Follow-up experiments validated regulatory relationships between genetically interacting pairs (Cdc28-Stb1 and Pho85-Pdr1), suggesting that GI studies involving OE alleles of regulatory proteins will be a rich source of new functional information.
authors.affiliation: Department of Molecular Genetics, University of Toronto, Ontario M5S 3E1, Canada.|Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
authors.forename: Ji-Young
authors.identifier: 0000-0001-6799-5709
authors.initials: JY
authors.lastname: Youn
authors.affiliation: Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
authors.forename: Helena
authors.identifier: 0000-0001-8925-7440
authors.initials: H
authors.lastname: Friesen
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Ontario M5S 3B2, Canada.
authors.forename: Alex N
authors.identifier: 0000-0003-1357-6386
authors.initials: AN
authors.lastname: Nguyen Ba
authors.affiliation: Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
authors.forename: Wendy
authors.identifier: 0000-0001-7454-8456
authors.initials: W
authors.lastname: Liang
authors.affiliation: Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
authors.forename: Vincent
authors.identifier: 0000-0003-0476-5425
authors.initials: V
authors.lastname: Messier
authors.affiliation: Department of Molecular Genetics, University of Toronto, Ontario M5S 3E1, Canada.|Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
authors.forename: Mike J
authors.initials: MJ
authors.lastname: Cox
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Ontario M5S 3B2, Canada.|Centre for the Analysis of Genome Evolution and Function, University of Toronto, Ontario M5S 3B2, Canada.
authors.forename: Alan M
authors.initials: AM
authors.lastname: Moses
authors.affiliation: Department of Molecular Genetics, University of Toronto, Ontario M5S 3E1, Canada brenda.andrews@utoronto.ca.|Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
authors.forename: Brenda
authors.identifier: 0000-0001-6427-6493
authors.initials: B
authors.lastname: Andrews
mesh_terms: D000483:Alleles; D018628:Gene Dosage; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D011494:Protein Kinases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D011494:Protein Kinases
keywords: genetic interactions; genetic networks; kinase; transcription factor; yeast genetics