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Sequence-Targeted Nucleosome Sliding in vivo - Nucleosome Mapping

ABSTRACT: Nucleosome positions were determined in wild type cells, cells lacking Isw2 or Ume6, and cells containing a hybrid Chd1-Ume6 chimeric remodeler

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE72570 | GEO | 2016/03/12

SECONDARY ACCESSION(S): PRJNA294376

REPOSITORIES: GEO

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Sequence-Targeted Nucleosome Sliding in vivo - Nucleosome Mapping

Project description:Nucleosome positions were determined in wild type cells, cells lacking Isw2 or Ume6, and cells containing a hybrid Chd1-Ume6 chimeric remodeler Matched MNase digests from W303 strain variants during log growth (OD600=0.4-0.6) were subject to paired-end sequencing for nucleosome mapping. For effects of the engineered fusion remodeling protein, a catalytically inactive (ATPase dead D513N) variant was included as a control.

2016-03-12 | E-GEOD-72570 | biostudies-arrayexpress

Stepwise reconstitution of genomic nucleosome organization with pure proteins

Project description:Nucleosome arrays begin at nucleosome-free promoter regions (NFRs) and regulate gene expression. Reconstituting such organization throughout a genome with purified proteins is a critical challenge in establishing biochemical mechanisms for chromosome assembly. Here we establish a four-step hierarchical building plan for yeast genomic nucleosome organization using only purified components: genomic DNA, histones, site-specific organizing factors Abf1 and Reb1, and the chromatin remodelers RSC, ISW2, INO80, and ISW1a. First, RSC makes NFRs by translating promoter poly(dA:dT) tracts into directional nucleosome removal. Second, +1 nucleosomes are positioned by INO80 at most genes potentially involving DNA shape, or by ISW2 using gene-specific Abf1 and Reb1. Third, INO80 or ISW2 create arrays with wide spacing. Fourth, ISW1a tightens the spacing and creates properly positioned arrays. We conclude that entire genomes use a simple set of rules and proteins, without transcription, to build a common chromatin architecture. In this study, nucleosomes were assembled using Salt Gradient Dialysis (SGD) on yeast genomic DNA library. Assembled nucleosomes were either left untreated (labelled as "SGD", control), treated with whole cell extract (WCE), mutant extracts (rsc3ts WCE, isw1 isw2 chd1 WCE), purified remodelers; singly or in combinations (RSC, ISW1a, ISW1b, ISW2, INO80, CHD1, SWI/SNF), combinations of mutant extracts and chromatin remodelers or combination of General Regulatory Factors (Abf1, Reb1) and chromatin remodelers. The resulting nucleosome positions were mapped genome-wide using MNase-(anti-H3-ChIP)-Seq.

2016-08-14 | E-GEOD-72106 | biostudies-arrayexpress

Sequence-Targeted Nucleosome Sliding in vivo

Project description:This SuperSeries is composed of the SubSeries listed below.

2016-03-12 | GSE72572 | GEO

Sequence-Targeted Nucleosome Sliding in vivo

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2016-03-12 | E-GEOD-72572 | biostudies-arrayexpress

The ISW1 and CHD1 ATP-dependent chromatin remodelers compete to set nucleosome spacing in vivo

Project description:We addressed the roles of three nucleosome spacing enzymes (ISW1, ISW2 and CHD1) in specifying chromatin organization in S. cerevisiae. Comparison of budding yeast nucleosome positions obtained using micrococcal nuclease digestion followed by paired-end sequencing (MNase-seq). We sequenced nucleosomes from isogenic strains with the isw1Î, isw2Î or chd1Î mutations in all combinations. We measured gene activity by ChIP-seq for the Rpb3 subunit of RNA polymerase II for some of these mutants.

2016-02-17 | E-GEOD-69400 | biostudies-arrayexpress

Nucleosome sliding by the Chd1 chromatin remodeler relies on the integrity of the DNA duplex

Project description:The experiments are done with three libraries by introducing base-pair perturbations on Widom 601: (1) poly(dA:dT) tract library (2) mismatch library (3) insertion library. And additional two more libraries based on native yeast genomic sequences: (4) Park97 mismatch library (5) Park97 insertion library. And we measured the nucleosome positioining in the base-pair resolution for before and after sliding by Chd1 chromatin remodeler.

2022-05-12 | GSE198440 | GEO

Sequence-Targeted Nucleosome Sliding in vivo - Transcription Profiling

Project description:RNA sequencing was performed on various W303 variants to determine effects of nucleosome repositioning on transcript abundance

2016-03-12 | GSE72571 | GEO

Sequence-Targeted Nucleosome Sliding in vivo - Transcription Profiling

Project description:RNA sequencing was performed on various W303 variants to determine effects of nucleosome repositioning on transcript abundance RNA sequencing was carried out in multiple backgrounds to determine effects of nucleosome repositioning in various contexts. For engineered chromatin remodeling factors, a catalytically inactive control was included.

2016-03-12 | E-GEOD-72571 | biostudies-arrayexpress

Isw2 and Ume6 ChIP-chip in transcription factor deletion (M-NM-^Tume6, M-NM-^Tnrg1, M-NM-^Tcin5, and M-NM-^Tsok2) and DNA-looping deficient (sua7-1) cells

Project description:The sequence-specific transcription factors Ume6, Nrg1, Cin5, and Sok2 and the general transcription factor TFIIB mediate the genome-wide targeting of the ATP-dependent chromatin remodeling enzyme Isw2. At least two biological replicates comparing Isw2-ChIP (Isw2-K215R ChIP DNA competitively hybridized against WT Isw2 ChIP DNA) in WT and M-NM-^Tume6, M-NM-^Tnrg1, M-NM-^Tcin5, M-NM-^Tsok2, and sua7-1 strains or Ume6-ChIP (ChIP DNA competitively hybridized again Input DNA) in WT and sua7-1 strains

2013-03-14 | E-GEOD-39542 | biostudies-arrayexpress

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

Project description:Chromatin remodelers are ATP-dependent enzymes that reorganize nucleosomes within all eukaryotic genomes. The Chd1 remodeler specializes in shifting nucleosomes into evenly spaced arrays, a defining characteristic of chromatin in gene bodies that blocks spurious transcription initiation. Linked to some forms of autism and commonly mutated in prostate cancer, Chd1 is essential for maintaining pluripotency in stem cells. Here we report a complex of yeast Chd1 bound to a nucleosome in a nucleotide-free state, determined by cryo-electron microscopy (cryo-EM) to 2.6 Å resolution. The structure shows a bulge of the DNA tracking strand where the ATPase motor engages the nucleosome, consistent with an initial stage in DNA translocation. Unlike other remodeler-nucleosome complexes, nucleosomal DNA compensates for the remodeler-induced bulge with a bulge of the complementary DNA strand one helical turn downstream from the ATPase motor. Unexpectedly, the structure also reveals an N-terminal binding motif, called ChEx, which binds on the exit-side acidic patch of the nucleosome. The ChEx motif can displace a LANA-based peptide from the acidic patch, which suggests a means by which Chd1 remodelers may block competing chromatin remodelers from acting on the opposite side of the nucleosome.

2022-02-23 | PXD025287 | Pride

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