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G3 (Bethesda). 2017 Mar 10;7(3):911-921. doi: 10.1534/g3.116.038471.

Functional Analysis of Kinases and Transcription Factors in Saccharomyces cerevisiae Using an Integrated Overexpression Library.

Author information

1
Department of Molecular Genetics, University of Toronto, Ontario M5S 3E1, Canada.
2
Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Ontario M5S 3E1, Canada.
3
Department of Cell and Systems Biology, University of Toronto, Ontario M5S 3B2, Canada.
4
Centre for the Analysis of Genome Evolution and Function, University of Toronto, Ontario M5S 3B2, Canada.
5
Department of Molecular Genetics, University of Toronto, Ontario M5S 3E1, Canada brenda.andrews@utoronto.ca.

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 cerevisiae We 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.

KEYWORDS:

genetic interactions; genetic networks; kinase; transcription factor; yeast genetics

PMID:
28122947
PMCID:
PMC5345721
DOI:
10.1534/g3.116.038471
[Indexed for MEDLINE]
Free PMC Article

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