title: Disruption of Drosophila Rad50 causes pupal lethality, the accumulation of DNA double-strand breaks and the induction of apoptosis in third instar larvae.
abstract: The Rad50/Mre11/Nbs1 protein complex has a crucial role in DNA metabolism, in particular in double-strand break (DSB) repair through homologous recombination (HR). To elucidate the role of the Rad50 protein complex in DSB repair in a multicellular eukaryote, we generated a Rad50 deficient Drosophila strain by P-element mediated mutagenesis. Disruption of Rad50 causes retarded development and pupal lethality. To investigate the mechanism of pupal death, brains and wing imaginal discs from third instar larvae were studied in more detail. Wing imaginal discs from Rad50 mutant larvae displayed a 3.5-fold increase in the induction of spontaneous apoptotic cells in comparison to their heterozygous siblings. This finding correlates with increased levels of phosphorylated histone H2Av, indicating an accumulation of DSBs in Rad50 mutant larvae. A 45-fold increase in the frequency of anaphase bridges was detected in the brains of Rad50 deficient larvae, consistent with a role for Rad50 in telomere maintenance and/or replication of DNA. The induction of DSBs and defects in chromosome segregation are in agreement with a role of Drosophila Rad50 in repairing the DSBs that arise during replication.
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands.
authors.forename: Marcin M
authors.initials: MM
authors.lastname: Gorski
authors.forename: Ron J
authors.initials: RJ
authors.lastname: Romeijn
authors.forename: Jan C J
authors.initials: JC
authors.lastname: Eeken
authors.forename: Anja W M
authors.initials: AW
authors.lastname: de Jong
authors.forename: Bert L
authors.initials: BL
authors.lastname: van Veen
authors.forename: Karoly
authors.initials: K
authors.lastname: Szuhai
authors.forename: Leon H
authors.initials: LH
authors.lastname: Mullenders
authors.forename: Wouter
authors.initials: W
authors.lastname: Ferro
authors.forename: Albert
authors.initials: A
authors.lastname: Pastink
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D001921:Brain / Q000502:physiology; D004247:DNA / Q000235:genetics*; D004249:DNA Damage; D045643:DNA Repair Enzymes; D004261:DNA Replication; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins / Q000502:physiology*; D004331:Drosophila melanogaster / Q000235:genetics*; D004706:Endodeoxyribonucleases / Q000502:physiology*; D029968:Escherichia coli Proteins; D005090:Exodeoxyribonucleases / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005804:Genes, Lethal; D006579:Heterozygote; D006657:Histones / Q000378:metabolism; D007814:Larva / Q000254:growth & development / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017386:Sequence Homology, Amino Acid; D014921:Wings, Animal / Q000502:physiology
chemical_list: C101253:DDP3 protein, E coli; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D029968:Escherichia coli Proteins; D006657:Histones; D004247:DNA; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases; C492905:Rad50 protein, Drosophila; D045643:DNA Repair Enzymes