The location and structure of double-strand DNA breaks induced during yeast meiosis: evidence for a covalently linked DNA-protein intermediate

EMBO J. 1995 Sep 15;14(18):4599-608.

Abstract

We have determined the precise location and structure of the double-strand DNA breaks (DSBs) formed during Saccharomyces cerevisiae meiosis. Breaks were examined at two recombination hot spots in both wild-type and rad50S mutant cells. At both loci, breaks occurred at multiple, irregularly spaced sites in a approximately 150 nucleotide interval contained within an area of nuclease-hypersensitive chromatin. No consensus sequence could be discerned at or around break sites. Patterns of cleavage observed on individual strands indicated that breaks initially form with a two nucleotide 5' overhang. Broken strands from rad50S mutant cells contained tightly bound protein at their 5' ends. We suggest that, in S.cerevisiae, meiotic recombination is initiated by a DSB-forming activity that creates a covalently linked protein-DNA intermediate.

MeSH terms

  • Argininosuccinate Lyase
  • Base Sequence
  • Chromatin / chemistry
  • Chromosome Mapping
  • DNA Damage*
  • DNA Repair / genetics
  • DNA, Fungal / genetics*
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins*
  • Fungal Proteins / analysis
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Meiosis / genetics*
  • Molecular Sequence Data
  • Promoter Regions, Genetic / genetics
  • Protein Binding
  • Recombination, Genetic*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Sequence Analysis, DNA

Substances

  • Chromatin
  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • RAD50 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Arg4 protein, S cerevisiae
  • Argininosuccinate Lyase