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Exploring Quantitative Yeast Phenomics with Single-Cell Analysis of DNA Damage Foci
Erin B Styles1, Karen J Founk1, Lee A Zamparo2
1The Donnelly Centre, University of Toronto, Toronto, ON M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 3E1, Canada.
Cell Systems
|September 13, 2016
Summary
This study introduces a new high-content screening method for functional genomics, enabling genome-wide analysis of DNA damage foci in yeast mutants. The approach identified novel genes involved in DNA repair and protein interactions.
Area of Science:
- Cell Biology
- Genomics
- Biochemistry
Background:
- Functional genomics requires scalable methods for analyzing cell biological data at the genome scale.
- Understanding sub-cellular structures and morphology is crucial for deciphering cellular processes.
Purpose of the Study:
- To develop an integrated method combining genome-wide genetic perturbation and high-content screening for yeast.
- To genetically map sub-cellular structures and compartment morphology using a DNA damage focus assay.
Main Methods:
- Genome-wide genetic perturbation in Saccharomyces cerevisiae.
- High-content screening using a Rad52-GFP marker for DNA damage foci.
- Machine learning-based automated image analysis for phenotype classification.
Main Results:
- Examined ~20 million cells across ~5,000 mutant backgrounds under genetic/chemical perturbations.
- Identified 345 mutants with elevated DNA damage foci, many dependent on sensitized backgrounds.
- Discovered Vid22 interacts with Sgs1 helicase at DNA damage sites, binding G-quadruplex regions.
Conclusions:
- The integrated method provides a scalable approach for functional genomics and cell biology.
- This method is extensible to various cell biological markers and experimental systems.
- Vid22's role in DNA damage response and G-quadruplex binding was elucidated.
