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Hrp3-TAP_Purification from Schizosaccharomyces pombe

ABSTRACT: Hrp3_Purification from Schizosaccharomyces pombe 972h- Eukaryotic genome is composed of repeating units of nucleosomes to form chromatin arrays. A canonical gene is marked by nucleosome free region (NFR) at its 5’ end followed by uniformly spaced arrays of nucleosomes. In fission yeast we show both biochemically and in vivo that both Hrp1 and Hrp3 are key determinants of uniform spacing of genic arrays.

OTHER RELATED OMICS DATASETS IN: PRJNA177319

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Schizosaccharomyces Pombe 972h-

SUBMITTER: Ignasi Forne

LAB HEAD: Ignasi Forne

PROVIDER: PXD000041 | Pride | 2014-05-16

REPOSITORIES: Pride

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			Action	DRS
	F030392.dat	Other
	F030393.dat	Other
	F030394.dat	Other
	F030395.dat	Other
	F030396.dat	Other

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Publications

CHD1 remodelers regulate nucleosome spacing in vitro and align nucleosomal arrays over gene coding regions in S. pombe.

Pointner Julia J Persson Jenna J Prasad Punit P Norman-Axelsson Ulrika U Strålfors Annelie A Khorosjutina Olga O Krietenstein Nils N Svensson J Peter JP Ekwall Karl K Korber Philipp P

The EMBO journal 20121026 23

Nucleosome positioning governs access to eukaryotic genomes. Many genes show a stereotypic organisation at their 5'end: a nucleosome free region just upstream of the transcription start site (TSS) followed by a regular nucleosomal array over the coding region. The determinants for this pattern are unclear, but nucleosome remodelers are likely critical. Here we study the role of remodelers in global nucleosome positioning in S. pombe and the corresponding changes in expression. We find a striking ...[more]

PMID: 23103765

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Insights into promoter chromatin remodeling from the structure of ISW1a complex bound to the dinucleosome

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MNase-seq analysis of Col-0 and chr11-1 chr17-1

Project description:Chromatin has a highly organized structure with the repeating nucleosome subunits. The position of nucleosomes on the chromatin is dynamically regulated by ATP-dependent chromatin remodeling factors (remodelers), therefore providing specific epigenetic information. However, the in vivo nucleosome distribution pattern in plants and how plant remodelers control the pattern formation are not yet clear. Here we use the Micrococcal Nuclease digestion followed by deep sequencing (MNase-seq) assay to show the genome-wide nucleosome pattern in plants and the role of the Arabidopsis thaliana Imitation Switch (AtISWI) subfamily remodelers in the pattern formation. Our data revealed three patterns in wild-type plants: 1) Promoters have a relative low nucleosome density compared with gene bodies; 2) Nucleosome density is negatively associated with gene expression levels; 3) The evenly and periodically spaced nucleosomes in gene bodies are positively associated with gene expression levels. Double mutations in the AtISWI genes CHROMATIN REMODELING 11 (CHR11) and CHR17 resulted in loss of the evenly-spaced-nucleosome pattern in gene bodies, but did not affect nucleosome density, suggesting that the primary role of AtISWI is to slide nucleosomes in gene bodies for promotion of transcription. The overall design of the experiment: (1) Nuclear extraction from Col-0 and chr11-1 chr17-1. (2) MNase treatment. (3) DNA purification. (4) Library construction. (5) Deep sequencing. (6) Data analysis.

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MNase-seq analysis of Col-0 and chr11-1 chr17-1

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Nucleosome recognition and DNA distortion by the Chd1 remodeler revealed at 2.6 Å resolution

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