A Novel Approach for Identifying Synthetic Dosage Lethal Interactions in Pooled Yeast Cultures

• Main Author: Ralph, Alison Carly
• Other Authors: Andrews, Brenda
• Language: en_ca
• Published: 2012
• Subjects: yeast; genomics; kinases; genetics; barcode; overexpression; 0369; 0307
• Online Access: http://hdl.handle.net/1807/33726

Systematic genomic studies in the budding yeast S. cerevisiae have greatly increased our capacity to conduct functional profiling of the eukaryotic genome. I describe a new method that makes use of yeast “Barcoder” strains to uniquely tag strains in a yeast overexpression collection (FLEX) and to systematically examine the effects of gene overexpression on cell fitness. This novel system is compatible with the so-called Synthetic Genetic Array (SGA) method, which automates yeast genetics and allows introduction of marked query alleles of interest into arrayed collections of yeast mutants. I identified SDL interactions for two key regulatory kinases, Dun1 and Mck1, using my system. I also used my array and approach to identify SDL interactions for Dun1 that are only manifest in the presence of DNA damage. These studies demonstrate the utility of the pooled SGA-SDL method for systematic discovery of condition-specific genetic interactions in conserved biological pathways.