A Phenomic Assessment of Subcellular Morphology in Saccharomyces cerevisiae Using Synthetic Genetic Array Analysis and High-Content Screening

In this thesis, I describe the optimization and adaptation of an integrated SGA-HCS pipeline complete with a robust panel of downstream statistical metrics, to analyze subcellular phenotypes in yeast cells in the context of the yeast deletion collection (Giaever et al., 2002a; Winzeler, 1999) and the ts collection of essential gene mutants (Li et al., 2011a). I have used this experimental pipeline to closely examine the utility of both single and double mutant screening strategies for the identification of subtle subcellular phenotypes, with a special focus on the formation of DNA damage foci, and have subsequently characterized Vid22, a protein with a previously unappreciated role in the DNA damage response. I have additionally endeavored to curate known cellular and subcellular mutant morphologies into a database, using a systematic rhetoric to describe conditions and observable phenotypes in order to provide a resource and gold standard set of mutants with which to assess ongoing Marker Project screens.