title: Polynucleotide kinase/phosphatase, Pnk1, is involved in base excision repair in Schizosaccharomyces pombe.
abstract: We previously reported that Schizosaccharomyces pombe pnk1 cells are more sensitive than wild-type cells to γ-radiation and camptothecin, indicating that Pnk1 is required for DNA repair. Here, we report that pnk1pku70 and pnk1rhp51 double mutants are more sensitive to γ-radiation than single mutants, from which we infer that Pnk1's primary role is independent of either homologous recombination or non-homologous end joining mechanisms. We also report that pnk1 cells are more sensitive than wild-type cells to oxidizing and alkylating agents, suggesting that Pnk1 is involved in base excision repair. Mutational analysis of Pnk1 revealed that the DNA 3'-phosphatase activity is necessary for repair of DNA damage, whereas the 5'-kinase activity is dispensable. A role for Pnk1 in base excision repair is supported by genetic analyses which revealed that pnk1apn2 is synthetically lethal, suggesting that Pnk1 and Apn2 may function in parallel pathways essential for the repair of endogenous DNA damage. Furthermore, the nth1pnk1apn2 and tdp1pnk1apn2 triple mutants are viable, implying that single-strand breaks with 3'-blocked termini produced by Nth1 and Tdp1 contribute to synthetic lethality. We also examined the sensitivity to methyl methanesulfonate of all single and double mutant combinations of nth1, apn2, tdp1 and pnk1. Together, our results support a model where Tdp1 and Pnk1 act in concert in an Apn2-independent base excision repair pathway to repair 3'-blocked termini produced by Nth1; and they also provide evidence that Pnk1 has additional roles in base excision repair.
authors: Department of Biochemistry & Molecular Biology, University of Calgary, Calgary, AB T2N4N1, Canada.
authors: Ekaterina
authors: E
authors: Kashkina
authors: Tao
authors: T
authors: Qi
authors: Michael
authors: M
authors: Weinfeld
authors: Dallan
authors: D
authors: Young
mesh_terms: D002166:Camptothecin / Q000494:pharmacology; D053903:DNA Breaks, Double-Stranded; D053904:DNA Breaks, Single-Stranded; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics*; D004271:DNA, Fungal / Q000187:drug effects / Q000528:radiation effects; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000235:genetics / Q000378:metabolism; D005720:Gamma Rays / Q000009:adverse effects; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation; D009708:Nucleotidases / Q000235:genetics / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000235:genetics / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004271:DNA, Fungal; D009097:Multienzyme Complexes; D009153:Mutagens; C492078:Nth1 protein, S pombe; C084232:RHP51 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; C539712:Tdp1 protein, S pombe; D008741:Methyl Methanesulfonate; D011494:Protein Kinases; C121377:protein kinase U; C448179:PnK1 protein, S pombe; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D051135:Rad51 Recombinase; D009708:Nucleotidases; D010727:Phosphoric Diester Hydrolases; D045647:DNA Glycosylases; C480729:Apn2 protein, S pombe; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D002166:Camptothecin