A double-strand break repair component is essential for S phase completion in fission yeast cell cycling

Mol Biol Cell. 1999 Oct;10(10):3331-43. doi: 10.1091/mbc.10.10.3331.

Abstract

Fission yeast rad22(+), a homologue of budding yeast RAD52, encodes a double-strand break repair component, which is dispensable for proliferation. We, however, have recently obtained a cell division cycle mutant with a temperature-sensitive allele of rad22(+), designated rad22-H6, which resulted from a point mutation in the conserved coding sequence leading to one amino acid alteration. We have subsequently isolated rad22(+) and its novel homologue rti1(+) as multicopy suppressors of this mutant. rti1(+) suppresses all the defects of cells lacking rad22(+). Mating type switch-inactive heterothallic cells lacking either rad22(+) or rti1(+) are viable, but those lacking both genes are inviable and arrest proliferation with a cell division cycle phenotype. At the nonpermissive temperature, a synchronous culture of rad22-H6 cells performs DNA synthesis without delay and arrests with chromosomes seemingly intact and replication completed and with a high level of tyrosine-phosphorylated Cdc2. However, rad22-H6 cells show a typical S phase arrest phenotype if combined with the rad1-1 checkpoint mutation. rad22(+) genetically interacts with rad11(+), which encodes the large subunit of replication protein A. Deletion of rad22(+)/rti1(+) or the presence of rad22-H6 mutation decreases the restriction temperature of rad11-A1 cells by 4-6 degrees C and leads to cell cycle arrest with chromosomes incompletely replicated. Thus, in fission yeast a double-strand break repair component is required for a certain step of chromosome replication unlinked to repair, partly via interacting with replication protein A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Bleomycin / pharmacology
  • Cell Cycle / genetics
  • Cell Division / genetics
  • Cloning, Molecular
  • DNA Repair / genetics*
  • DNA-Binding Proteins / genetics
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics*
  • Gene Expression Regulation, Fungal
  • Genes, Fungal*
  • Genes, Mating Type, Fungal*
  • Molecular Sequence Data
  • Mutation
  • Replication Protein A
  • S Phase / genetics
  • Schizosaccharomyces pombe Proteins*
  • Suppression, Genetic
  • Temperature
  • Yeasts / genetics*

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • Replication Protein A
  • Schizosaccharomyces pombe Proteins
  • rad52 protein, S pombe
  • Bleomycin