The location and structure of double-strand DNA breaks induced during yeast meiosis: evidence for a covalently linked DNA-protein intermediate.
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.
Liu
J
J
Laboratory of Biochemistry, National Cancer Institute, Bethesda, MD 20892-4255, USA.
Wu
T C
TC
Lichten
M
M
D001123:Argininosuccinate Lyase; D001483:Base Sequence; D002843:Chromatin / Q000737:chemistry; D002874:Chromosome Mapping; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008540:Meiosis / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA
D002843:Chromatin; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C086930:RAD50 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C087900:Arg4 protein, S cerevisiae; D001123:Argininosuccinate Lyase