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Replication origin mapping of mitotic and meiotic cells in different conditions

ABSTRACT: Identification of sites of replication initiation by copy number analysis, comparing samples before and after entry into S phase. Experiments were performed in the following strains and conditions for mitotic cells: cdc25-22 (temperature-sensitive mutation arrests cells at G2/M for synchrony, otherwise wild type), nitrogen-rich conditions; wild type, haploid and diploid, nitrogen-depletion conditions. Experiments were performed in the following strains and conditions for meiotic cells: pat1-114 diploid (temperature-sensitive mutation induces a synchronous meiosis, otherwise wild type), nitrogen-depletion and nitrogen-rich conditions; pat1-114 diploid with altered Cdc45 levels, nitrogen-depletion conditions.

ORGANISM(S): Schizosaccharomyces pombe

PROVIDER: GSE53879 | GEO | 2014/02/20

SECONDARY ACCESSION(S): PRJNA233629

REPOSITORIES: GEO

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Replication origin mapping of mitotic and meiotic cells in different conditions

Project description:Identification of sites of replication initiation by copy number analysis, comparing samples before and after entry into S phase. Experiments were performed in the following strains and conditions for mitotic cells: cdc25-22 (temperature-sensitive mutation arrests cells at G2/M for synchrony, otherwise wild type), nitrogen-rich conditions; wild type, haploid and diploid, nitrogen-depletion conditions. Experiments were performed in the following strains and conditions for meiotic cells: pat1-114 diploid (temperature-sensitive mutation induces a synchronous meiosis, otherwise wild type), nitrogen-depletion and nitrogen-rich conditions; pat1-114 diploid with altered Cdc45 levels, nitrogen-depletion conditions. Comparison of samples before and after S phase entry; dye-swap experiments were performed and averaged.

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DNA intermediates of meiotic recombination in synchronous S. pombe at optimal temperature

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Transcription profiling of fission yeast meiosis and sporulation

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Transcription profiling of S. pombe wild type and meu5delta cells during pat1 induced meiosis, wild type meiosis and overexpression of the mei4 transcription factor

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DNA intermediates of meiotic recombination in synchronous S. pombe at optimal temperature

Project 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.

2013-09-20 | GSE49961 | GEO

Transcription profiling of fission yeast meiosis and sporulation

Project description:pat1 meiosis is very synchronous (see E-SNGR-2), but the use of a temperature shift and the prolonged nitrogen starvation can lead to artifacts. As a control for the E-SNGR-2 experiment, non-sporulating pat1+ diploids were treated exactly as in the protocol for E-SNGR-2 cells. Under these conditions the cells re-enter the mitotic cycle. As a reference we used a pool consisting of equal amount of RNA from all time points of the experiments. This experiment is part of a series, E-SNGR-2 to E-SNGR-7.

2002-08-05 | E-SNGR-5 | biostudies-arrayexpress

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