title: Genetic Regulation of Dna2 Localization During the DNA Damage Response.
abstract: DNA damage response pathways are crucial for protecting genome stability in all eukaryotes. Saccharomyces cerevisiae Dna2 has both helicase and nuclease activities that are essential for Okazaki fragment maturation, and Dna2 is involved in long-range DNA end resection at double-strand breaks. Dna2 forms nuclear foci in response to DNA replication stress and to double-strand breaks. We find that Dna2-GFP focus formation occurs mainly during S phase in unperturbed cells. Dna2 colocalizes in nuclear foci with 25 DNA repair proteins that define recombination repair centers in response to phleomycin-induced DNA damage. To systematically identify genes that affect Dna2 focus formation, we crossed Dna2-GFP into 4293 nonessential gene deletion mutants and assessed Dna2-GFP nuclear focus formation after phleomycin treatment. We identified 37 gene deletions that affect Dna2-GFP focus formation, 12 with fewer foci and 25 with increased foci. Together these data comprise a useful resource for understanding Dna2 regulation in response to DNA damage.
authors.affiliation: Department of Biochemistry and Donnelly Centre, University of Toronto, Toronto, ON M5S 3E1, Canada.
authors.forename: Askar
authors.initials: A
authors.lastname: Yimit
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195.
authors.forename: Michael
authors.initials: M
authors.lastname: Riffle
authors.affiliation: Department of Biochemistry and Donnelly Centre, University of Toronto, Toronto, ON M5S 3E1, Canada grant.brown@utoronto.ca.
authors.forename: Grant W
authors.identifier: http://orcid.org/0000-0002-9002-5003
authors.initials: GW
authors.lastname: Brown
mesh_terms: D053903:DNA Breaks, Double-Stranded; D004249:DNA Damage; D004265:DNA Helicases / Q000378:metabolism*; D004260:DNA Repair; D016195:G2 Phase; D015966:Gene Expression Regulation, Fungal; D042822:Genomic Instability; D011485:Protein Binding; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D021381:Protein Transport; D051721:Rad52 DNA Repair and Recombination Protein / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: D051721:Rad52 DNA Repair and Recombination Protein; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D004265:DNA Helicases; C410222:DNA2 protein, S cerevisiae
keywords: DNA damage; DNA repair; Dna2 foci; Saccharomyces cerevisiae; colocalization; double-strand breaks; genetics; genome stability; nuclear foci