title: Spindle Checkpoint Factors Bub1 and Bub2 Promote DNA Double-Strand Break Repair by Nonhomologous End Joining.
abstract: The nonhomologous end-joining (NHEJ) pathway is essential for the preservation of genome integrity, as it efficiently repairs DNA double-strand breaks (DSBs). Previous biochemical and genetic investigations have indicated that, despite the importance of this pathway, the entire complement of genes regulating NHEJ remains unknown. To address this, we employed a plasmid-based NHEJ DNA repair screen in budding yeast (Saccharomyces cerevisiae) using 369 putative nonessential DNA repair-related components as queries. Among the newly identified genes associated with NHEJ deficiency upon disruption are two spindle assembly checkpoint kinases, Bub1 and Bub2. Both observation of resulting phenotypes and chromatin immunoprecipitation demonstrated that Bub1 and -2, either alone or in combination with cell cycle regulators, are recruited near the DSB, where phosphorylated Rad53 or H2A accumulates. Large-scale proteomic analysis of Bub kinases phosphorylated in response to DNA damage identified previously unknown kinase substrates on Tel1 S/T-Q sites. Moreover, Bub1 NHEJ function appears to be conserved in mammalian cells. 53BP1, which influences DSB repair by NHEJ, colocalizes with human BUB1 and is recruited to the break sites. Thus, while Bub is not a core component of NHEJ machinery, our data support its dual role in mitotic exit and promotion of NHEJ repair in yeast and mammals.
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada Department of Biology and Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada.
authors.forename: Matthew
authors.initials: M
authors.lastname: Jessulat
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: Ramy H
authors.initials: RH
authors.lastname: Malty
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: Diem-Hang
authors.initials: DH
authors.lastname: Nguyen-Tran
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: Viktor
authors.initials: V
authors.lastname: Deineko
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aoki
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: James
authors.initials: J
authors.lastname: Vlasblom
authors.affiliation: Department of Biology and Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada.
authors.forename: Katayoun
authors.initials: K
authors.lastname: Omidi
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada Banting and Best Department of Medical Research, Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Ke
authors.initials: K
authors.lastname: Jin
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: Zoran
authors.initials: Z
authors.lastname: Minic
authors.affiliation: Department of Biology and Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Hooshyar
authors.affiliation: Department of Biology and Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Burnside
authors.affiliation: Department of Biology and Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada.
authors.forename: Bahram
authors.initials: B
authors.lastname: Samanfar
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
authors.forename: Sadhna
authors.initials: S
authors.lastname: Phanse
authors.affiliation: Cancer Research Unit, Saskatchewan Cancer Agency, Saskatoon, Saskatchewan, Canada.
authors.forename: Tanya
authors.initials: T
authors.lastname: Freywald
authors.affiliation: Department of Medicine, Regina Qu'Appelle Health Region, Regina, Saskatchewan, Canada.
authors.forename: Bhanu
authors.initials: B
authors.lastname: Prasad
authors.affiliation: Banting and Best Department of Medical Research, Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Zhaolei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Cancer Research Unit, Saskatchewan Cancer Agency, Saskatoon, Saskatchewan, Canada.
authors.forename: Franco
authors.initials: F
authors.lastname: Vizeacoumar
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.
authors.forename: Nevan J
authors.initials: NJ
authors.lastname: Krogan
authors.affiliation: Cancer Research Unit, Saskatchewan Cancer Agency, Saskatoon, Saskatchewan, Canada.
authors.forename: Andrew
authors.initials: A
authors.lastname: Freywald
authors.affiliation: Department of Biology and Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada ashkan_golshani@carleton.ca mohan.babu@uregina.ca.
authors.forename: Ashkan
authors.initials: A
authors.lastname: Golshani
authors.affiliation: Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada ashkan_golshani@carleton.ca mohan.babu@uregina.ca.
authors.forename: Mohan
authors.initials: M
authors.lastname: Babu
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D064447:Checkpoint Kinase 2 / Q000235:genetics / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D004260:DNA Repair; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000235:genetics; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D000071857:Tumor Suppressor p53-Binding Protein 1
chemical_list: C070351:BUB2 protein, S cerevisiae; C584280:Bub1 protein, S cerevisiae; D018797:Cell Cycle Proteins; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D029701:Saccharomyces cerevisiae Proteins; C498301:TP53BP1 protein, human; D000071857:Tumor Suppressor p53-Binding Protein 1; D064447:Checkpoint Kinase 2; C584277:BUB1 protein, human; D017346:Protein Serine-Threonine Kinases; C067022:RAD53 protein, S cerevisiae