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Whole-genome nucleosome mapping in meiotic diploid S. cerevisiae

ABSTRACT: We determined nucleosome positions throughout the genome in diploid S. cerevisiae undergoing early stages of synchronous meiosis. This study sought to assess if systematic reorganization of nucleosomes occurs during meiotic prophase at or near sites of DNA double strand break formation. Six samples were sequenced, each a separate time point from either of two meiotic time courses. Dataset N1 consists of samples collected at 0, 1, 2, and 3 hr in meiosis; Dataset N2 consists of samples collected at 0 and 4 hr. Time courses, sample preparation, and sequencing were carried out in separate laboratories for the two datasets: Hochwagen lab (Whitehead Institute) for Dataset N1; Keeney lab (Howard Hughes Medical Institute; Memorial Sloan-Kettering Cancer Center) for Dataset N2.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Scott keeney

PROVIDER: E-GEOD-26452 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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A hierarchical combination of factors shapes the genome-wide topography of yeast meiotic recombination initiation.

Pan Jing J Sasaki Mariko M Kniewel Ryan R Murakami Hajime H Blitzblau Hannah G HG Tischfield Sam E SE Zhu Xuan X Neale Matthew J MJ Jasin Maria M Socci Nicholas D ND Hochwagen Andreas A Keeney Scott S

Cell 20110301 5

The nonrandom distribution of meiotic recombination influences patterns of inheritance and genome evolution, but chromosomal features governing this distribution are poorly understood. Formation of the DNA double-strand breaks (DSBs) that initiate recombination results in the accumulation of Spo11 protein covalently bound to small DNA fragments. By sequencing these fragments, we uncover a genome-wide DSB map of unprecedented resolution and sensitivity. We use this map to explore how DSB distribu ...[more]

PMID: 21376234

Similar Datasets

Genome-wide mapping of meiotic double-strand breaks by sequencing Spo11 oligos.

Project description:The nonrandom distribution of meiotic recombination shapes patterns of inheritance and genome evolution, but chromosomal features governing this distribution are poorly understood. Formation of the DNA double-strand breaks (DSBs) that initiate recombination results in accumulation of Spo11 protein covalently bound to small DNA fragments. We show here that sequencing these fragments provides a genome-wide DSB map of unprecedented resolution and sensitivity. We use this map to explore the influence of large-scale chromosome structures, chromatin, transcription factors, and local sequence composition on DSB distributions. Our analysis supports the view that the recombination terrain is molded by combinatorial and hierarchical interaction of factors that work on widely different size scales. Mechanistic aspects of DSB formation and early processing steps are also uncovered. This map illuminates the occurrence of DSBs in repetitive DNA elements, repair of which can lead to chromosomal rearrangements. We discuss implications for evolutionary dynamics of recombination hotspots. We deep sequenced 4 samples of Spo11 oligos on Roche 454 platform. Three samples are technical replicates of Spo11 oligo products prepared from one meiotic culture and the fourth sample was prepared from an independent culture.

2011-03-07 | E-GEOD-26449 | biostudies-arrayexpress

Chromatin immunoprecipitation to determine binding of ARS binding factor 1 (Abf1) in yeast during vegetative growth and meiotic development

Project description:Binding of the yeast ARS binding factor 1 (Abf1) to target promotors during vegetative growth and meiotic development as tested by ChIP-CHIP analysis;

2008-05-01 | E-MEXP-1439 | biostudies-arrayexpress

Transcription profiling by array of human CD4 positive cells after activation with antiCD3/28 beads

Project description:The positioning of nucleosomes with respect to DNA plays an important role in regulating transcription. However, nucleosome mapping has been performed for only limited genomic regions in humans. We have generated genome-wide maps of nucleosome positions in both resting and activated human CD4+ T cells by direct sequencing of nucleosome ends using the Solexa high-throughput sequencing technique. Corresponding ChIP-Seq data are available in NCBI's Short Read Archive at ftp://ftp.ncbi.nih.gov/pub/TraceDB/ShortRead/SRA000234/; We find that nucleosome phasing relative to the transcription start sites (TSSs) is directly correlated to RNA polymerase II binding. Furthermore, the first nucleosome downstream of TSSs exhibits differential positioning in active and silent genes. TCR signaling induces extensive nucleosome reorganization in promoters and enhancers to allow transcriptional activation or repression. Our results suggest that H2A.Z-containing and modified nucleosomes are preferentially lost from the -1 nucleosome position. Our data provide a comprehensive view of the nucleosome landscape and its dynamic regulation in the human genome. This microarray dataset is the gene expression data from resting and activated CD4+ T cells. We used this data to look at nucleosome positioning, Pol II and a few chromatin modifications at genes that are silent and activated in both resting and activated T cells as well as genes that are induced or repressed with T cell activation. Experiment Overall Design: Gene expression data from resting (M0 and T0) and activated (M18 and T18) T cells.

2008-04-08 | E-GEOD-10437 | biostudies-arrayexpress

Translational repression of the Drosophila nanos mRNA involves the RNA helicase Belle and RNA coating by Me31B and Trailer hitch

Project description:The maternal mRNA nanos is required for anterior-posterior patterning in the early Drosophila melanogaster development. The nanos-mRNA is translationally repressed and deadenylated during the first two hours of embryogenesis. A stem-loop structure in the 3'-UTR of this RNA (SRE - Smaug recogniiton element) is bound by the Smaug protein which recruits a protein complex to the RNA to sequester its translation and intiate transcript degradation via the ccr4/not-complex. To analyse the composition of this repressor complex, biotinylated RNAs carrying two SRE stem-loops were used for an affinity purification of bound proteins. As a negative control an RNA was used that contained two point mutations in the SRE loop, that abolish binding of Smaug. Seven subunits were identified as components of the repressor complex. The main deadenylase in Drosophila, the ccr4/not-complex was also significantly enriched.

2017-07-18 | PXD006596 | Pride

Comparative genome-wide DNA methylation analysis of colorectal tumor and matched normal tissues

Project description:In our study we applied a genome-wide DNA methylation analysis approach, MethylCap-seq, to map the differentially methylated regions in 24 tumor and matched normal colon samples. In total, 2687 frequently hypermethylated and 468 frequently hypomethylated regions were identified, which include potential biomarkers for CRC diagnosis. Hypermethylation in the tumor samples was enriched at CpG islands and gene promoters, while hypomethylation was distributed throughout the genome. Using epigenetic data from human embryonic stem cells, we show that frequent differentially methylated regions (DMRs) coincide with bivalent loci in human embryonic stem cells. DNA methylation is commonly thought to lead to cancer gene related silencing, however integration of publically available expression analysis shows that 75% of the frequently hypermethylation genes were most likely already lowly or not expressed in normal tissue. Collectively, our study provides genome-wide DNA methylation maps of colon cancer, comprehensive lists of DMRs, and gives further clues on the role of aberrant DNA methylation in CRC formation. To investigate DNA methylation in CRC in a genome-wide unbiased fashion, we applied MethylCap-seq. This method involves capture of methylated DNA using the MBD domain of MeCP2, and subsequent next-generation Illumina sequencing of eluted DNA. In addition, we compared MethylCap with RNA-seq and ChIP-seq profiles of H3K4me3 and H3K27me3 for the colon cancer tumor cell line HCT116 (HCT116 WT) and the cell line of HCT116 with DNMT1 and DNMT3b knockout (HCT116 DKO).

2012-10-26 | E-GEOD-39068 | biostudies-arrayexpress

scATAC-seq of mouse Plasmodium-specific CD4+ T cells

Project description:Plasmodium-specific CD4+ T cells from mice infected with Plasmodium chabaudi chabaudi AS parasites were recovered at Days 0, 4, 7, and 32 to undergo processing and to generate scATAC-seq dataset. At Day 7, CXCR5+ and CXCR6+ cells were recovered separately. At Day 32, mice were administered with either saline or artesunate (intermittent artesunate therapy - IAT). scATAC-seq dataset was analysed to investigate epigenomic landscapes of CD4+ T cells from effector to memory states.

2020-10-14 | E-MTAB-9402 | biostudies-arrayexpress

Rec114 ChIP-seq

Project description:DNA duplication is intimately connected to setting up post-replicative chromosome structures and events, but molecular details of this coordination are not well understood. A striking example occurs during yeast meiosis, where replication locally influences timing of the DNA double-strand breaks (DSBs) that initiate recombination. We show here that replication-DSB coordination is eliminated by overexpressing Dbf4-dependent Cdc7 kinase (DDK) or removing Tof1 or Csm3, components of the replication fork protection complex (FPC). DDK physically associates with Tof1, and Tof1 is dispensable for replication-DSB coordination if DDK is artificially tethered to replisomes. Furthermore, DDK phosphorylation of the DSB-promoting factor Mer2 is locally coordinated with replication, dependent on Tof1. These findings indicate that DDK recruited by FPC to replisomes phosphorylates chromatin-bound Mer2 in the wake of the replication fork, thus synchronizing replication with an early prerequisite for DSB formation. This may be a general mechanism to ensure spatial and temporal coordination of replication with other chromosomal processes. Ninety-six samples total: 12 time points (each time points contains ChIP and input samples) from Rec114-myc ARS+, Rec114-myc arsM-bM-^HM-^F strains, Rec114-myc tof1M-bM-^HM-^FARS+ and Rec114-myc tof1M-bM-^HM-^F arsM-bM-^HM-^F strains

2014-07-21 | E-GEOD-52970 | biostudies-arrayexpress

Mapping of cohesion factor association sites in S. cerevisiae - Meiosis I chromosome segregation is established through regulation of microtubule-kinetochore interactions.

Project description:During meiosis, a single round of DNA replication is followed by two consecutive rounds of nuclear divisions called meiosis I and meiosis II. In meiosis I, homologous chromosomes segregate, while sister chromatids remain together. Determining how this unusual chromosome segregation behavior is established is central to understanding germ cell development. Here we show that preventing microtubule-kinetochore interactions during premeiotic S phase and prophase I is essential for establishing the meiosis I chromosome segregation pattern. Premature interactions of kinetochores with microtubules transform meiosis I into a mitosis-like division by disrupting two key meiosis I events: coorientation of sister kinetochores and protection of centromeric cohesin removal from chromosomes. Furthermore we find that restricting outer kinetochore assembly contributes to preventing premature engagement of microtubules with kinetochores. We propose that inhibition of microtubule-kinetochore interactions during premeiotic S phase and prophase I is central to establishing the unique meiosis I chromosome segregation pattern. The association of the cohesion factors Rec8, Pds5, and Sgo1 were measured by ChIP-chip analysis in wild-type and CUP-CLB3 strains.

2012-10-05 | E-GEOD-41339 | biostudies-arrayexpress

Pytheas SARS-CoV-2 spike protein mRNA sample analysis

Project description:Mass spectrometry remains an important method for analysis of modified nucleosides ubiquitously present in cellular RNAs, in particular for ribosomal and transfer RNAs that play crucial roles in mRNA translation and decoding. Furthermore, modifications have effect on the lifetimes of nucleic acids in plasma and cells and are consequently incorporated into RNA therapeutics. To provide an analytical tool for sequence characterization of modified RNAs, we developed Pytheas, an open-source software package for automated analysis of tandem MS data for RNA. This dataset contains the analysis of 14N and 15N-labeled synthetic mRNA of the SARS-CoV-2 spike protein. All uridines were modified to m1Ψ.

2022-05-04 | PXD030845 | Pride

Polygenic and directional regulatory evolution across pathways in Saccharomyces

Project description:We assess the prevalence of polygenic evolution in pathways between the yeasts S. cerevisiae and S. bayanus. We first established short-read sequencing methods to detect cis-regulatory variation in a diploid hybrid between the species. We then formulated an analytic strategy to test for the scenario in which selective pressure in one species, to increase or decrease the activity of a pathway, has driven the accumulation of cis-regulatory variants that act in the same direction on gene expression. Application of this test revealed a variety of yeast pathways with evidence for directional regulatory evolution. Measurement of allele-specific expression of each ortholog in a diploid hybrid between Saccharomyces species

2010-02-22 | E-GEOD-19837 | biostudies-arrayexpress

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