title: Msh1p counteracts oxidative lesion-induced instability of mtDNA and stimulates mitochondrial recombination in Saccharomyces cerevisiae.
abstract: The proximity of the mitochondrial genome to the respiratory chain, a major source of ROS (radical oxygen species), makes mtDNA more vulnerable to oxidative damage than nuclear DNA. Mitochondrial BER (base excision repair) is generally considered to be the main pathway involved in the prevention of oxidative lesion-induced mutations in mtDNA. However, we previously demonstrated that the increased frequency of mitochondrial Oli(r) mutants in an ogg1Delta strain, lacking the activity of a crucial mtBER glycosylase, is reduced in the presence of plasmids encoding Msh1p, the mitochondrial homologue of the bacterial mismatch protein MutS. This finding suggested that Msh1p might be involved in the prevention of mitochondrial mutagenesis induced by oxidative stress. Here we show that a double mutant carrying the msh1-R813W allele, encoding a variant of the protein defective in the ATP hydrolysis activity, combined with deletion of SOD2, encoding the mitochondrial superoxide dismutase, displays a synergistic effect on the frequency of Oli(r) mutants, indicating that Msh1p prevents generation of oxidative lesion-induced mitochondrial mutations. We also show that double mutants carrying the msh1-R813W allele, combined with deletion of either OGG1 or APN1, the latter resulting in deficiency of the Apn1 endonuclease, exhibit a synergistic effect on the frequency of respiration-defective mutants having gross rearrangements of the mitochondrial genome. This suggests that Msh1p, Ogg1p and Apn1p play overlapping functions in maintaining the stability of mtDNA. In addition, we demonstrate, using a novel ARG8(m) recombination assay, that a surplus of Msh1p results in enhanced mitochondrial recombination. Interestingly, the mutant forms of the protein, msh1p-R813W and msh1p-G776D, fail to stimulate recombination. We postulate that the Msh1p-enhanced homologous recombination may play an important role in the prevention of oxidative lesion-induced rearrangements of the mitochondrial genome.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland. anetak@ibb.waw.pl
authors.forename: Aneta
authors.initials: A
authors.lastname: Kaniak
authors.forename: Piotr
authors.initials: P
authors.lastname: Dzierzbicki
authors.forename: Agata T
authors.initials: AT
authors.lastname: Rogowska
authors.forename: Ewa
authors.initials: E
authors.lastname: Malc
authors.forename: Marta
authors.initials: M
authors.lastname: Fikus
authors.forename: Zygmunt
authors.initials: Z
authors.lastname: Ciesla
mesh_terms: D045647:DNA Glycosylases / Q000172:deficiency; D045643:DNA Repair Enzymes / Q000172:deficiency; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000172:deficiency; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases / Q000172:deficiency; D005656:Fungal Proteins / Q000502:physiology*; D042822:Genomic Instability; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism; D024101:Mitochondrial Proteins; D009154:Mutation; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000235:genetics*; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D013482:Superoxide Dismutase / Q000172:deficiency; D000099156:Superoxide Dismutase 2
chemical_list: D045647:DNA Glycosylases; D045643:DNA Repair Enzymes; D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D005656:Fungal Proteins; D006147:Guanine; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2; C078660:MSH1 protein, S cerevisiae; C024829:8-hydroxyguanine; C083274:Apn1 protein, S cerevisiae; C505160:Ogg1 protein, S cerevisiae; C101825:NTG1 protein, S cerevisiae