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Genome-wide binding of the meiosis specific Rec8 cohesin subunit in metaphase I

ABSTRACT: ChIP-seq profile of Rec8 binding sites arrested in metaphase I by depletion of the APC/C activator CDC20. The aim of this experiment is to identify Rec8 binding sites in metaphase I.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE123546 | GEO | 2019/04/25

REPOSITORIES: GEO

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Genome-wide binding of the cohesin protector Sgo1 in the presence and absence of the meiosis I-specific Spo13 protein

Project description:ChIP-seq profile of Sgo1 binding sites arrested in metaphase I by deletion of the APC/C activator CDC20. The aim of this experiment is to identify whether Sgo1 binding differs in wild type cells and cells deficient of the meiosis I-specific protein Spo13, which is required for retention of pericentromeric cohesin after metaphase I.

2019-04-25 | GSE112167 | GEO

Genome-wide binding of the meiosis specific Rec8 cohesin subunit in metaphase I

Project description:Genome-wide binding of the meiosis specific Rec8 cohesin subunit in metaphase I

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Proteomics profiling of mitotic arrest by transcriptional silencing of the Cdc20 homolog slp1 in fission yeast

Project description:Depletion of the APC/C activator Cdc20 arrests cells in metaphase with high levels of the mitotic cyclin (Cyclin B) and the Separase inhibitor Securin. In mammalian cells this arrest has been exploited for the treatment of cancer with drugs that engage the spindle assembly checkpoint and, recently, with chemical inhibitors of the APC/C. While most cells arrested in mitosis for prolonged periods undergo apoptosis, others skip cytokinesis and enter G1 with unsegregated chromosomes. This process, known as mitotic slippage, generates aneuploidy and increases genomic instability in the cancer cell. Here, we analyse the behaviour of fission yeast cells arrested in mitosis through the transcriptional silencing of the Cdc20 homolog slp1. While depletion of slp1 readily halts cells in metaphase, this arrest is only transient and a majority of cells eventually undergo cytokinesis and show steady mitotic dephosphorylation. As a result, we generated this label-free dataset with the aim of providing further insights into the global dynamics of protein expression under slp1 depletion and the underlying mechanisms that contribute to the mitotic escape.

2022-10-13 | PXD031975 | Pride

Pmk1 kinase-dependent phosphosite mapping on Slp1

Project description:In late mitosis, the timely polyubiquitylation and subsequent degradation of securin and cyclin B by anaphase-promoting complex/cyclosome (APC/C) play a critical role for anaphase onset and chromosome segregation. Binding of Cdc20, the co-activator of the APC/C, is critical for activation of APC/C at anaphase entry. We found that one MAPK, Pmk1, is involved in phosphorylation of Slp1Cdc20 (the Cdc20 homologue in the fission yeast Schizosaccharomyces pombe), which may promote its ubiquitylation and subsequent degradation, and thus impede APC/C activation and anaphase entry. To gain a full picture of Pmk1-dependent phosphosite map of Slp1Cdc20, we purified the APC/C-associated Slp1Cdc20 by immunoprecipitation of one APC/C subunit Apc15 in metaphase-arrested nda3-km311 cells. Apc15-GFP was purified from wild type, pek1DD [i.e. pek1(S234D,T238D)] and pmk1∆ cells respectively. Samples were separated by SDS-PAGE, and Coomassie blue-stained gel slices were excised and submitted for mass spectrometry analyses.

2024-10-15 | PXD048800 | Pride

DNA double-strand breaks regulate the cleavage-independent release of Rec8-cohesin during yeast meiosis

Project description:The mitotic cohesin complex necessary for sister chromatid cohesion and chromatin loop formation shows local and global association to chromosomes in response to DNA double-strand breaks (DSBs). Here, by genome-wide binding analysis of the meiotic cohesin with Rec8 as a kleisin, we found that Rec8 shows dynamic localization from middle to late meiotic prophase I with cleavage-independent global dissociation along chromosomes, driven by meiotic DSB formation. Each Rec8 binding site on the chromosome axis follows distinct dynamics with dissociation and association. Centromeres also showed reduced Rec8 binding in late prophase I relative to mid-prophase I, implying chromosome remodeling of the regions. Rec8 dissociation profile per chromosome is largely correlated with meiotic DSB density. Indeed, the spo11 mutant deficient in meiotic DSB formation did not show the cleavage-independent dissociation of Rec8 in late meiotic prophase I. These suggest the presence of a regulatory pathway for global Rec8-cohesin dynamics in response to meiotic DSBs.

2023-12-31 | GSE224074 | GEO

Genome-wide binding of the meiosis I-specific Spo13 in the presence and absence of cohesin

Project description:ChIP-seq profile of Spo13 binding sites arrested in prophase I by deletion of the meiotic transcription factor NDT80. Previous ChIP-ChIP data had indicated that Spo13 binding correlates with cohesin binding sites. Therefore, in addition to wild type cells, we also investigated cells deficient of the meiosis-specific cohesin subunit Rec8 to identify cohesin-dependent binding sites of Spo13.

2019-04-25 | GSE112170 | GEO

Calibrated ChIP-seq for meiotic-specific cohesin (Rec8) and mitotic-specific cohesin (Scc1) in budding yeast cells arrested in metaphase of mitosis

Project description:Calibrated ChIP-seq for meiotic-specific cohesin (Rec8) and mitotic-specific cohesin (Scc1) in budding yeast cells arrested in metaphase of mitosis

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Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling

Project description:Accurate chromosome segregation is coordinated by the spindle assembly checkpoint (SAC) through its effector the mitotic checkpoint complex (MCC), to inhibit the anaphase-promoting complex or cyclosome (APC/C). Cdc20 is an essential mitotic regulator since it promotes mitotic exit through activating the APC/C and monitors kinetochore-microtubule attachment through activating the SAC. The proper functioning of Cdc20 requires multiple interactions with APC/C and MCC subunits. To functionally assess each of these interactions within cells requires efficient depletion of endogenous Cdc20, which is highly difficult to achieve by RNAi. Here we generated Cdc20 RNAi sensitive cell lines by CRISPR/Cas9 which display a penetrant metaphase arrest phenotype by a single RNAi treatment. In this null background, we accurately measured the contribution of each known motif of Cdc20 on APC/C and SAC activation. The CRY box, a previously identified degron was found to be critical for the SAC by promoting theMCC formation and stabilizing the interaction between the MCC and APC/C. These data reveal additionalregulatory components within the SAC and establish a novel method to interrogate Cdc20 function.

2023-12-06 | PXD045411 | Pride

Genome wide map of DMC1 in testis of Stag3-/-, Rec8-/- and Stag3-/-Rec8-/- mice.

Project description:During mouse meiosis, DNA double-strand breaks (DSBs) are initiated by SPO11 at recombination hotspots (HSs), activated by PRDM9. Although activated HSs are marked by H3K4me3 and H3K36me3 histone modifications at open chromatin, most of the DSB-initiating and repair proteins are associated with the chromosome axis. This study addresses the mechanistic importance of the axis-associated cohesin proteins in DSB formation. We demonstrate that interactions between PRDM9 and the meiotic axis proteins STAG3 and REC8 are essential for efficient DSB formation. The absence of STAG3 or REC8 leads to inefficient meiotic DSB formation at HSs. STAG3 is critical for DSB formation, even at PRDM9-independent DSB-sites. Also, STAG3 and REC8 facilitate recruitment of the DSB-promoting proteins HORMAD1, IHO1 and MEI4 required for SPO11 activity. Together, these results support an evolutionarily conserved model in which axis-associated cohesin complexes recruit recombination-initiating proteins to DSB sites to promote meiotic recombination initiation.

2019-03-07 | GSE112110 | GEO

Chip-seq analysis of meiotic Cohesin (mei-Cohesins) subunit REC8 cleaved by separase (REC8*), and SMC3 in Macaca fascicularis testis

Project description:The study aimed to map epigenomic locations of separase-cleaved REC8 using chromatin IP analyzed by high-throughput sequencing on purified germ cells from M. fascicularis testis

2022-11-07 | GSE201989 | GEO

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