title: Dissecting DNA damage response pathways by analysing protein localization and abundance changes during DNA replication stress.
abstract: Relocalization of proteins is a hallmark of the DNA damage response. We use high-throughput microscopic screening of the yeast GFP fusion collection to develop a systems-level view of protein reorganization following drug-induced DNA replication stress. Changes in protein localization and abundance reveal drug-specific patterns of functional enrichments. Classification of proteins by subcellular destination enables the identification of pathways that respond to replication stress. We analysed pairwise combinations of GFP fusions and gene deletion mutants to define and order two previously unknown DNA damage responses. In the first, Cmr1 forms subnuclear foci that are regulated by the histone deacetylase Hos2 and are distinct from the typical Rad52 repair foci. In a second example, we find that the checkpoint kinases Mec1/Tel1 and the translation regulator Asc1 regulate P-body formation. This method identifies response pathways that were not detected in genetic and protein interaction screens, and can be readily applied to any form of chemical or genetic stress to reveal cellular response pathways.
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors: Johnny M
authors: JM
authors: Tkach
authors: Askar
authors: A
authors: Yimit
authors: Anna Y
authors: AY
authors: Lee
authors: Michael
authors: M
authors: Riffle
authors: Michael
authors: M
authors: Costanzo
authors: Daniel
authors: D
authors: Jaschob
authors: Jason A
authors: JA
authors: Hendry
authors: Jiongwen
authors: J
authors: Ou
authors: Jason
authors: J
authors: Moffat
authors: Charles
authors: C
authors: Boone
authors: Trisha N
authors: TN
authors: Davis
authors: Corey
authors: C
authors: Nislow
authors: Grant W
authors: GW
authors: Brown
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D004249:DNA Damage; D004261:DNA Replication / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D017353:Gene Deletion; D006655:Histone Deacetylases / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D021381:Protein Transport / Q000235:genetics / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: C111825:ASC1 protein, S cerevisiae; D048868:Adaptor Proteins, Signal Transducing; C573119:Cmr1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D029701:Saccharomyces cerevisiae Proteins; C088937:MEC1 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases; C513623:TEL1 protein, S cerevisiae; D006655:Histone Deacetylases; D019204:GTP-Binding Proteins