Tkach JM~Brown GW, 2012

Pubmed ID 22842922
Title Dissecting DNA damage response pathways by analysing protein localization and abundance changes during DNA replication stress.
Authors Johnny M Tkach, Askar Yimit, Anna Y Lee, Michael Riffle, Michael Costanzo, Daniel Jaschob, Jason A Hendry, Jiongwen Ou, Jason Moffat, Charles Boone, Trisha N Davis, Corey Nislow, Grant W Brown
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.
Citation Nat. Cell Biol. 2012; 14:966-76


Download the list of datasets
Paper Phenotype Condition Medium Collection Tested mutants Data Details
Tkach JM~Brown GW, 2012 P-body formation (Lsm1-GFP) standard SD + His + Leu + Met + Ura, ammonium sulphate hap a N/A None
Tkach JM~Brown GW, 2012 P-body formation (Lsm1-GFP) hydroxyurea [200 mM] SD + His + Leu + Met + Ura, ammonium sulphate hap a N/A None

Curation history

Tested strains

Feb. 3, 2018 To request.
Feb. 7, 2019 Request sent.


Feb. 3, 2018 To request.
Feb. 7, 2019 Request sent.