title: Fission yeast homologs of human XPC and CSB, rhp41 and rhp26, are involved in transcription-coupled repair of methyl methanesulfonate-induced DNA damage.
abstract: Methyl methanesulfonate (MMS) methylates nitrogen atoms in purines, and predominantly produces 7-methylguanine and 3-methyladenine (3-meA). Previously, we showed that base excision repair (BER) and nucleotide excision repair (NER) synergistically function to repair MMS-induced DNA damage in the fission yeast Schizosaccharomyces pombe. Here, we studied the roles of NER components in repair of 3-meA and BER intermediates such as the AP site and single strand breaks. Mutants of rhp41 (XPC homolog) and rhp26 (CSB homolog) exhibited moderate sensitivity to MMS. Transcription of the fbp1 gene, which is induced by glucose starvation, was strongly inhibited by MMS damage in rhp41Δ and rhp26Δ strains but not in wild type and 3-meA DNA glycosylase-deficient cells. The results indicate that Rhp41p and Rhp26p are involved in transcription-coupled repair (TCR) of MMS-induced DNA damage. In the BER pathway of S. pombe, AP lyase activity of Nth1p mainly incises the AP site to generate a 3'-blocked end, which is in turn converted to 3'-OH by Apn2p. Deletion of rad16 or rhp26 in the nth1Δ strain greatly enhanced MMS sensitivity, suggesting that the AP site could also be corrected by TCR. Double mutant apn2Δ/rad16Δ exhibited hypersensitivity to MMS, implying that Rad16p provides a backup pathway for removal of the 3'-blocked end. Moreover, an rhp51Δ strain was extremely sensitive to MMS and double mutants of nth1Δ/rhp51Δ and apn2Δ/rhp51Δ increased the sensitivity, suggesting that homologous recombination is necessary for repair of three different types of lesions, 3-meA, AP sites and 3'-blocked ends.
authors.affiliation: Department of Biochemistry, Faculty of Science, Okayama University of Science, Okayama 700-0005, Japan.
authors.forename: Kyoichiro
authors.initials: K
authors.lastname: Kanamitsu
authors.forename: Shogo
authors.initials: S
authors.lastname: Ikeda
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism; D053904:DNA Breaks, Single-Stranded; D004265:DNA Helicases / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004260:DNA Repair / Q000235:genetics / Q000502:physiology*; D045643:DNA Repair Enzymes / Q000235:genetics; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D015966:Gene Expression Regulation, Fungal / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008741:Methyl Methanesulfonate; D009154:Mutation / Q000235:genetics; D000075223:Poly-ADP-Ribose Binding Proteins; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000502:physiology*; D016019:Survival Analysis
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; C101945:RHP16 protein, S pombe; C482447:Rhp41 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; C025946:3-methyladenine; D008741:Methyl Methanesulfonate; D004265:DNA Helicases; C078673:ERCC6 protein, human; C480729:Apn2 protein, S pombe; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D045643:DNA Repair Enzymes; D000225:Adenine