title: Nonsense-mediated decay regulates key components of homologous recombination.
abstract: Cells frequently experience DNA damage that requires repair by homologous recombination (HR). Proteins involved in HR are carefully coordinated to ensure proper and efficient repair without interfering with normal cellular processes. In Saccharomyces cerevisiae, Rad55 functions in the early steps of HR and is regulated in response to DNA damage through phosphorylation by the Mec1 and Rad53 kinases of the DNA damage response. To further identify regulatory processes that target HR, we performed a high-throughput genetic interaction screen with RAD55 phosphorylation site mutants. Genes involved in the mRNA quality control process, nonsense-mediated decay (NMD), were found to genetically interact with rad55 phospho-site mutants. Further characterization revealed that RAD55 transcript and protein levels are regulated by NMD. Regulation of HR by NMD extends to multiple targets beyond RAD55, including RAD51, RAD54 and RAD57 Finally, we demonstrate that loss of NMD results in an increase in recombination rates and resistance to the DNA damaging agent methyl methanesulfonate, suggesting this pathway negatively regulates HR under normal growth conditions.
authors.affiliation: Department of Microbiology & Molecular Genetics, University of California, Davis, CA 95616-8665, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Janke
authors.affiliation: Department of Microbiology & Molecular Genetics, University of California, Davis, CA 95616-8665, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Kong
authors.affiliation: Department of Cellular & Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158-2517, USA.
authors.forename: Hannes
authors.initials: H
authors.lastname: Braberg
authors.affiliation: Department of Cell Biology, SR-11, Scripps Research institute, La Jolla, CA 92307, USA.
authors.forename: Greg
authors.initials: G
authors.lastname: Cantin
authors.affiliation: Department of Cell Biology, SR-11, Scripps Research institute, La Jolla, CA 92307, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.affiliation: Department of Cellular & Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158-2517, USA California Institute for Quantitative Biosciences, QB3, San Francisco, CA 94158-2517, USA J. David Gladstone Institute, San Francisco, CA, 94158-2517, USA.
authors.forename: Nevan J
authors.initials: NJ
authors.lastname: Krogan
authors.affiliation: Department of Microbiology & Molecular Genetics, University of California, Davis, CA 95616-8665, USA Department of Molecular & Cellular Biology University of California, Davis, CA 95616-8665, USA wdheyer@ucdavis.edu.
authors.forename: Wolf-Dietrich
authors.initials: WD
authors.lastname: Heyer
mesh_terms: D016000:Cluster Analysis; D019295:Computational Biology / Q000379:methods; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004843:Epistasis, Genetic; D015966:Gene Expression Regulation, Fungal; D059765:Homologous Recombination; D009154:Mutation; D059365:Nonsense Mediated mRNA Decay; D010766:Phosphorylation; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; C087191:RAD55 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins