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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Hrp3 controls nucleosome positioning to suppress non-coding transcription in eu- and heterochromatin

ABSTRACT: This SuperSeries is composed of the following subset Series: GSE37697: Antisense Transcripts in Mutant Strains GSE40451: Hrp3 controls nucleosome positioning to suppress non-coding transcription in eu- and heterochromatin [MNase-Seq] Refer to individual Series

ORGANISM(S): Schizosaccharomyces pombe

SUBMITTER: Yoonjung Choi

PROVIDER: E-GEOD-40453 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Hrp3 controls nucleosome positioning to suppress non-coding transcription in eu- and heterochromatin.

Shim Young Sam YS Choi Yoonjung Y Kang Keunsoo K Cho Kun K Oh Seunghee S Lee Junwoo J Grewal Shiv I S SI Lee Daeyoup D

The EMBO journal 20120918 23

The positioning of the nucleosome by ATP-dependent remodellers provides the fundamental chromatin environment for the regulation of diverse cellular processes acting on the underlying DNA. Recently, genome-wide nucleosome mapping has revealed more detailed information on the chromatin-remodelling factors. Here, we report that the Schizosaccharomyces pombe CHD remodeller, Hrp3, is a global regulator that drives proper nucleosome positioning and nucleosome stability. The loss of Hrp3 resulted in n ...[more]

PMID: 22990236

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Hrp3 controls nucleosome positioning to suppress non-coding transcription in eu- and heterochromatin [MNase-Seq]

Project description:We report that the Schizosaccharomyces pombe CHD remodeler, Hrp3, is a global regulator that drives higher-order chromatin structure and genomic stability. The loss of Hrp3 resulted in nucleosome perturbation across the chromosome, and the production of antisense transcripts in the hrp3M-bM-^HM-^F cells emphasized the importance of nucleosome architecture for proper transcription Examination of nucleosome positions in wild-type and mutants using the MNase-seq method

2012-10-02 | E-GEOD-40451 | biostudies-arrayexpress

Eic1 links Mis18 with the CCAN/Mis6/Ctf19 complex to promote CENP-A assembly

Project description:CENP-A chromatin the foundation for kinetochore assembly in all eukaryotes. CENP-A is incorporated outside of S phase and depends on its chaperone HJURPScm3, and Mis18 in vertebrates and fission yeast. The recruitment of Mis18 and HJURPScm3 to centromeres is cell cycle regulated. Vertebrate Mis18 associates with Mis18BP1KNL2 and RbAp46/48Mis16/Hat2. Mis18BP1KNL2 is critical for the recruitment of Mis18 and HJURPScm3 to vertebrate centromeres. However, no ortholog of Mis18BP1KNL2 has been identified in fission yeast, consequently it remains unknown how the key Cnp1CENP-A loading factor Mis18 is recruited. We identify two novel Mis18-interacting proteins (Eic1 and Eic2) in fission yeast. Our analyses show that Eic1 and Eic2 are components of a Mis18 complex. Eic1 is essential to maintain normal Cnp1CENP-A levels at centromeres and therefore is crucial for kinetochore integrity whereas Eic2 is dispensable. Eic1 also associates with components of the large constitutive CCAN/Sim4/Ctf19 complex. Our findings suggest that Eic1 is the functional counterpart of Mis18BP1KNL2 and acts as a link with constitutive kinetochore components to allow the temporally regulated recruitment of the Mis18 Cnp1CENP-A loader. 100bp paired end ChIP-Seq analysis of GFP tagged Eic1, Eic2, Scm3, and Mis18 with sonicated whole cell extract input DNA in asynchornous Schizosaccharomyces pombe cells.

2014-05-19 | E-GEOD-54685 | biostudies-arrayexpress

Sequence selectivity of macrolide-induced translational regulation

Project description:We sequenced a total of 12 cDNA libararies derived from fragmented total mRNA and ribosome protected mRNAs from azithromycin-treated and -untreated S. aureus samples. The data represented 2 independent biological replicates. Examination of the impact of azithromycin on global translatome, mRNA abundance and the ribosome density along the transcripts.

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Spt6 regulates intragenic and antisense transcription, nucleosome positioning, and histone modifications genome-wide in fission yeast [ChIP-seq]

Project description:Spt6 is a highly conserved histone chaperone that interacts directly with both RNA polymerase II and histones to regulate gene expression. To gain a comprehensive understanding of the requirements for this critical factor, we have performed genome-wide analyses of transcription, chromatin structure, and histone modifications in an S. pombe spt6 mutant. Our results demonstrate several dramatic changes to transcription and chromatin structure in the spt6 mutant, including an elevation of antisense transcripts at over 70 percent of all genes and general loss of the +1 nucleosome. Furthermore, Spt6 is required for the trimethylation of histone H3 on lysines 4 and 36, marks associated with active transcription. Taken together, our results indicate that Spt6 is critical for the accuracy of transcription and the integrity of chromatin, likely via its direct interactions with RNA polymerase II and histones. ChIP-seq experiments were performed on wild type and spt6-1 strains on the following proteins: RNA polymerase II (Rpb1), Paf1 Complex (Ctr9), COMPASS (Swd1), Set2, Spt6, histones H2B and H3, histone modifications H3K4me3 and H3K36me3. Experiments were performed in replicates and matching inputs were also sequenced.

2013-10-09 | E-GEOD-49574 | biostudies-arrayexpress

Antisense Transcripts in Mutant Strains

Project description:The positioning of the nucleosome by ATP-dependent nucleosome remodelers provides the fundamental chromatin environment for the regulation of diverse cellular processes acting on the underlying DNA. Here, we report that the fission yeast CHD remodeler, Hrp3, is a global regulator that drives higher-order chromatin structure and genomic stability. The loss of Hrp3 resulted in the production of antisense transcripts and perturbation of the nucleosome in an ATPase mutant of Hrp3 was also associated with destabilization of the DNA-histone interaction. Furthermore, the effect of Hrp3 in the pericentric region was found to be accomplished via a physical interaction with Swi6, and appeared to cooperate with other heterochromatin factors for gene silencing. Taken together, our data indicate that a well-positioned nucleosome is important for the spatial-temporal control of transcription-associated processes, and show that this is accomplished by the chromatin remodeler, Hrp3, in fission yeast. Two-condition experiment, Mutant vs Wild-type (control)

2012-05-07 | E-GEOD-37697 | biostudies-arrayexpress

RNAi mediates post-transcriptional repression of gene expression in fission yeast Schizosaccharomyces pombe [Affymetrix]

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2014-01-23 | E-GEOD-54190 | biostudies-arrayexpress

Fission yeast Cactin restricts telomere transcription and elongation by promoting Rap1 pre-mRNA splicing and protein stabilization

Project description:RNA transcriptome analysis of S.pombe fission yeast cells deleted for cay1+/SPBC2F12.12c (cay1D) and wildtype The transcriptome of cay1+ deleted fission yeast cells was characterized and compared to wildtype yeasts to identify differentially expressed genes controlled by Cay1 on a transcriptional or post-transcriptional level. 3 biological replicates of wildtype and cay1D fission yeast cells were prepared in parallel. Based on poor quality array hybridization one cay1D replicate was excluded from further analysis

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Project description: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.

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The ATP-dependent chromatin remodeling enzyme Fun30 represses transcription by sliding promoter proximal nucleosomes [seq]

Project description:The evolutionarily conserved ATP-dependent chromatin remodeling enzyme Fun30 has recently been shown to play important roles in heterochromatin silencing and DNA repair. However, how Fun30 remodels nucleosomes is not clear. Here we report a nucleosome sliding activity of Fun30 and its role in transcriptional repression. We observed that Fun30 repressed the expression of genes involved in amino acid and carbohydrate metabolism, the stress response, and meiosis. In addition, Fun30 was localized at the 5M-bM-^@M-2 and 3M-bM-^@M-2 ends of genes and within the open reading frames of its targets. Consistent with its role in gene repression, we observed that Fun30-target genes lacked histone modifications often associated with gene activation and showed an increased level of ubiquitinated histone H2B. Furthermore, genome-wide nucleosome mapping analysis revealed that the length of the nucleosome-free region at the 5M-bM-^@M-2 end of a subset of genes was changed in Fun30-depleted cells. In addition, the positions of the -1, +2 and +3 nucleosomes at the 5M-bM-^@M-2 end of target genes were significantly shifted, while position of the +1 nucleosome remained largely unchanged in the fun30M-NM-^T mutant. Finally, we demonstrated that affinity purified single-component Fun30 exhibited nucleosome sliding activity in an ATP-dependent manner. These results define a role for Fun30 in regulation of transcription and indicate that Fun30 remodels chromatin at the 5M-bM-^@M-2 end of genes by sliding promoter proximal nucleosomes. Micrococcal nuclease digested mononucleosome DNA from wild type and fun30D cells were sequenced by illumina Genome Analyzer. Genome-wide nucleosome positioning and occupancy were analyzed.

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