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

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

PROVIDER: PRJNA439308 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6871340_1.fastq.gz	Fastqsanger.gz
	SRR6871340_2.fastq.gz	Fastqsanger.gz
	SRR6871350_1.fastq.gz	Fastqsanger.gz
	SRR6871350_2.fastq.gz	Fastqsanger.gz
	SRR6871341_1.fastq.gz	Fastqsanger.gz

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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

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Project description:We report testis H3K4me3 and DMC1 enrichment at DSB hotspots in various mice to examine differences between infertile hybrid mice and mice that have been humanized at PRDM9, which have rescued fertility. We find that infertile mice have an excess of "asymmetric" DSB hotspots, where both H3K4me3 and DMC1 reads tend to originate from only one homologue. At these hotspots, we see an excess of DMC1 relative to H3K4me3, consistent with delayed DSB repair at these sites. See Davies et al. Nature 2016 for a complete summary.

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Genome wide maps of Dmc1 in testis of Hop2 null mice.

Project description:We report the application of ChIP-seq targeted at the meiosis-specific protein DMC1 to reveal the genome-wide distribution of initiation of meiotic recombination. The mouse model here employed is Hop2-/- because it is unable to repair the DNA double-stranded breaks and therefore the DMC1 signal is more persistent. We also provide the resulting dataset of ChIP-seq targeted at RAD51 which is not meiosis specific but is also targeted at initiation of recombination loci in meiotic tissue. In addition, we report DMC1 ChIP-seq on wild type mouse pup testis. Finally, we present ChIP-seq targeted at H3K4me3 in testis and liver tissues.

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ssDNA mapping in dmc1 strains

Project description:ssDNA enrichment was used to map and compare DSB hotspots in dmc1, pch2 dmc1, sir2 dmc1, orc1-161 dmc1, dmc1 rdnadelete and dmc1 chr2:12 translocation strains. 6 samples, 2 replicates each. Averaged data is available as a supplementary file on the Series record (below).

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Chromatin IP for Mcm2-7, Rec8, Hop1 and Red1

Project description:Mcm2-7 ChIP in pre-meiotic and pre-mitotic cells, axis factor ChIP in wild-type and replication compromised strains in meiosis Multiple studies of meiotic chromosomes were undertaken. To study DNA replication, the locations of replicative helicase (Mcm2-7) were mapped in pre-meiotic and pre-mitotic cells, and DNA replication profiles were created for pre-meiotic S (meiS) and pre-mitotic S (mitS) phases. Early origins were mapped in hydroxyurea for wild-type cells in mitS + 200mM HU, and meiS +20mM HU for wild-type, sml1, rec8 and spo11 deletion cells. Rec8, Hop1 and Red1 binding to meiotic chromosomes was evaluated using ChIP-chip in wild-type cells with and without 20 mM HU, and in cdc6-mn and clb5 clb6 delete cells. Finally, meiotic DNA double-strand breaks (DSBs) were mapped in cdc6-mn dmc1 delete cells by measuring the ssDNA that accumulates at DSB hotspots.

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Arabidopsis DMC1 ubiquitination sites identification, LC-MS/MS

Project description:To characterize the mechanism of DMC1 ubiquitination as well as further explore the impact of DMC1’s ubiquitination on meiosis, we attempted to identify the potential DMC1 ubiquitination sites. The in vitro ubiquitination assay using recombinant DMC1 heterologously expressed and purified from E. coli as substrate was conducted. Then mass spectrometry assay was performed to identify the ubiquitination sites of DMC1. Five DMC1 lysine residues (K45, K70, K101, K162, and K290) were identified as potential ubiquitination sites.

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