Youn JY, Friesen H, Nguyen Ba AN, Liang W, Messier V, Cox MJ, Moses AM, Andrews B

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 ...

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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 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 overexpression. We used the Synthetic Genetic Array (SGA) method to examine dosage-dependent genetic interactions (GIs) between 239 gain-of-function (overexpression) alleles of TFs and six loss-of-function and seven over-expression kinase alleles, the former identifying Synthetic Dosage Lethal (SDL) interactions and the latter testing a genetic interaction we call Double Dosage Lethality (DDL). We identified and confirmed 94 genetic interactions between 65 overexpression alleles of TFs and nine kinase alleles. Follow-up experiments validated regulatory relationships between genetically interacting pairs (Cdc28 - Stb1 and Pho85 - Pdr1), suggesting that genetic interaction studies involving overexpression alleles of regulatory proteins will be a rich source of new functional information.

Date: Jan. 25, 2017

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