E-GEOD-49961 - DNA intermediates of meiotic recombination in synchronous S. pombe at optimal temperature

Status
Released on 20 September 2013, last updated on 3 June 2014
Organism
Schizosaccharomyces pombe
Samples (12)
Array (1)
Protocols (6)
Description
Crossovers formed by recombination between homologous chromosomes are important for proper homolog segregation during meiosis and for generation of genetic diversity. Optimal molecular analysis of DNA intermediates of recombination requires synchronous cultures. We previously described a mutant, pat1-as2, of the fission yeast Schizosaccharomyces pombe that undergoes synchronous meiosis at 25°C when an ATP analog is added to the culture. Here, we compare recombination intermediates in pat1-as2 at 25°C with those in the widely used pat1-114 temperature-sensitive mutant at 34°C, a temperature higher than optimal. DNA double-strand breaks at most hotspots are similarly abundant in the two conditions but, remarkably, a few hotspots are distinctly deficient at 25°C. In both conditions Holliday junctions at DNA break hotspots form more frequently between sister chromatids than between homologs, but a novel species, perhaps arising from invasion by only one end of broken DNA, is more readily observed at 25°C. Our results confirm the validity of previous assays of recombination intermediates in S. pombe and provide new information on the mechanism of meiotic recombination. DNA double-strand break analysis by immunoprecipation of Rec12-FLAG covalently linked to DNA (without exogenous crosslinking agent used) following meiotic induction via pat1-114 or pat1-as2 alleles
Experiment type
ChIP-chip by tiling array 
Contacts
Kyle Fowler <krfowler@fhcrc.org>, Gerald R Smith, Juraj Gregan, Kyle R Fowler, Lubos Cipak, Randy W Hyppa
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-49961.idf.txt
Sample and data relationshipE-GEOD-49961.sdrf.txt
Raw data (1)E-GEOD-49961.raw.1.zip
Processed data (1)E-GEOD-49961.processed.1.zip
Array designA-GEOD-16421.adf.txt
Links