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Rec8 ChIP-seq for wild type, eco1-aa, wpl1Δ and eco1-aa wpl1Δ strains arrested in meiotic prophase I

ABSTRACT: Genome binding/occupancy profiling of the cohesin subunit Rec8 by high throughput sequencing.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE185018 | GEO | 2022/02/04

REPOSITORIES: GEO

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Calibrated Smc3 ChIP-seq for wild type, eco1-aa, wpl1-delta and eco1-aa wpl1-delta strains arrested in meiotic prophase I

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Rec8 ChIP-seq for wild type, eco1-aa, wpl1Δ and eco1-aa wpl1Δ strains arrested in meiotic prophase I

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Chromosome conformation for wild type, eco1-aa, cdc6-mn and eco1-aa cdc6-mn strains arrested in meiotic prophase I

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Meiotic cohesins modulate chromosome compaction during meiotic prophase in fission yeast

Project description:The meiotic cohesin Rec8 is required for the stepwise segregation of chromosomes during the two rounds of meiotic division. By directly measuring chromosome compaction in living cells of the fission yeast Schizosaccharomyces pombe, we found an additional role for the meiotic cohesin in the compaction of chromosomes during meiotic prophase. In the absence of Rec8, chromosomes were decompacted relative to those of wild-type cells. Conversely, loss of the cohesin-associated protein Pds5 resulted in hyper-compaction. While this hyper-compaction requires Rec8, binding of Rec8 to chromatin was reduced in the absence of Pds5, indicating that Pds5 promotes chromosome association of Rec8. To explain these observations, we propose that meiotic prophase chromosomes are organized as chromatin loops emanating from a Rec8-containing axis; the absence of Rec8 disrupts the axis, resulting in disorganized chromosomes, whereas reduced Rec8 loading results in a longitudinally compacted axis with fewer attachment points and longer chromatin loops. Keywords: ChIP-chip analysis ChIP analysis of Rec8: In all cases, hybridization data for ChIP fraction was compared with that of SUP (supernatant) fraction. Pombe chromosome II, III array was used.

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Rec8 ChIP-Seq experiment to analyse cohesin localisation in the Ctf19c mutants during meiotic prophase.

Project description:Ctf19c mutants were shown to specifically impair cohesin localisation at the pericentromere during mitosis. The goal of this experiment is to confirm whether this mutant has the same effect on cohesin localisation during meiotic prophase. To do so, diploid S. cerevisiae SK1 cells were constructed containing Rec8-3HA in an ndt80-d background. Cells were allowed to sporulate in sporulation media for 5 hours after which formaldehyde was added to the cultures and cells were fixed for 2hr. Samples were then processed according to the standard lab ChIP-Seq protocol (with 3 times 30cycles of 30sec of sonication) Diploid S. cerevisiae SK1 cells were constructed containing Rec8-3HA in an ndt80-d background. Cells were allowed to sporulate in sporulation media for 5 hours after which formaldehyde was added to the cultures and cells were fixed for 2hr. Samples were then processed according to the standard lab ChIP-Seq protocol (with 3 times 30cycles of 30sec of sonication)

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Meiotic cohesins modulate chromosome compaction during meiotic prophase in fission yeast

2006-08-23 | GSE5284 | GEO

Rec8 ChIP-Seq experiment to analyse cohesin localisation in the Ctf19c mutants during meiotic prophase.

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Rec8 cohesin-mediated loop-axis chromatin architecture is required for meiotic recombination

Project description:During meiotic prophase, cohesin-dependent axial structures are formed in the synaptonemal complex (SC). However, functional correlation between these structure formation and cohesion remains elusive. Here we examined formation of the cohesin-dependent axial structure in fission yeast, which forms atypical SCs composed of linear elements (LinEs) resembling the lateral elements of SC but lacking the central elements, and found that Rec8 cohesin is crucial for the formation of the loop-axis structure within the atypical SC. Furthermore, the Rec8-mediated loop-axis structure is formed in the absence of LinEs, and provides a structural platform for aligning homologous chromosomes. We also succeeded to identify a rec8 mutant that lost the ability of assembling the loop-axis structure without losing cohesion. Remarkably, this mutant showed defects in LinE assembly, resulting in great reduction of meiotic recombination. Collectively, our results demonstrate an essential role of the Rec8-dependent loop-axis structure in LinE assembly, facilitating meiotic recombination.

2022-04-27 | GSE192516 | GEO

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