Molecular characterization of the role of the Schizosaccharomyces pombe nip1+/ctp1+ gene in DNA double-strand break repair in association with the Mre11-Rad50-Nbs1 complex

Mol Cell Biol. 2008 Jun;28(11):3639-51. doi: 10.1128/MCB.01828-07. Epub 2008 Mar 31.

Abstract

The Schizosaccharomyces pombe nip1(+)/ctp1(+) gene was previously identified as an slr (synthetically lethal with rad2) mutant. Epistasis analysis indicated that Nip1/Ctp1 functions in Rhp51-dependent recombinational repair, together with the Rad32 (spMre11)-Rad50-Nbs1 complex, which plays important roles in the early steps of DNA double-strand break repair. Nip1/Ctp1 was phosphorylated in asynchronous, exponentially growing cells and further phosphorylated in response to bleomycin treatment. Overproduction of Nip1/Ctp1 suppressed the DNA repair defect of an nbs1-s10 mutant, which carries a mutation in the FHA phosphopeptide-binding domain of Nbs1, but not of an nbs1 null mutant. Meiotic DNA double-strand breaks accumulated in the nip1/ctp1 mutant. The DNA repair phenotypes and epistasis relationships of nip1/ctp1 are very similar to those of the Saccharomyces cerevisiae sae2/com1 mutant, suggesting that Nip1/Ctp1 is a functional homologue of Sae2/Com1, although the sequence similarity between the proteins is limited to the C-terminal region containing the RHR motif. We found that the RxxL and CxxC motifs are conserved in Schizosaccharomyces species and in vertebrate CtIP, originally identified as a cofactor of the transcriptional corepressor CtBP. However, these two motifs are not found in other fungi, including Saccharomyces and Aspergillus species. We propose that Nip1/Ctp1 is a functional counterpart of Sae2/Com1 and CtIP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Conserved Sequence
  • DNA Breaks, Double-Stranded
  • DNA Repair*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Epistasis, Genetic
  • Exodeoxyribonucleases
  • Meiosis / genetics
  • Molecular Sequence Data
  • Mutation
  • Phosphorylation
  • Protein Structure, Tertiary / genetics
  • Rad51 Recombinase / genetics
  • Rad51 Recombinase / metabolism
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism*

Substances

  • Chromosomal Proteins, Non-Histone
  • Ctp1 protein, S pombe
  • DNA-Binding Proteins
  • Nbs1 protein, S pombe
  • RHP51 protein, S pombe
  • Rad50 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • Rad51 Recombinase
  • Exodeoxyribonucleases
  • Mre11 protein, S pombe