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Series GSE35524 Query DataSets for GSE35524
Status Public on Nov 06, 2012
Title Mediator promotes CENP-A incorporation at fission yeast centromeres [ChIP-seq]
Organism Schizosaccharomyces pombe
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary At Schizosaccharomyces pombe centromeres, heterochromatin formation is required for de novo incorporation of the histone H3 variant CENP-A/Cnp1, which in turn directs kinetochore assembly and ultimately chromosome segregation during mitosis. Noncoding RNAs (ncRNAs) transcribed by RNA polymerase II (Pol II) directs heterochromatin formation via the RNAi machinery, but also through RNAiindependent RNA processing factors. Control of centromeric ncRNA transcription is therefore a key factor for proper centromere function. We here use transcriptional profiling, gene inactivation experiments, and chromatin immunoprecipitation analyses to demonstrate that the Mediator complex directs ncRNA transcription and regulates centromeric heterochromatin formation in fission yeast. Mediator co-localizes with Pol II at centromeres and loss of the Mediator subunit Med20 causes a dramatic increase in pericentromeric transcription and desilencing of the core centromere. As a consequence, heterochromatin formation is impaired both via the RNAi dependent and independent pathways, resulting in loss of CENP-A/Cnp1 from the core centromere, defect kinetochore function, and a severe chromosome segregation defect. Interestingly, the increased centromeric transcription observed in med20Δ appears to directly block CENP-A/Cnp1 incorporation and inhibition of Pol II transcription can suppress the observed phenotypes. Our data thus identify Mediator as a crucial regulator of ncRNA transcription at fission yeast centromeres and add another crucial layer of regulation to centromere function.
Overall design 3 samples examined: wild type chromatin incubated with beads as the non antibody control, wild type chromatin incubated with RNA Polymerase II CTD domain antibody and Protein G beads, and TAP-Med7 cells chromatin incubated with IgG beads.
Contributor(s) Carlsten JO, Zhu X, Davila Lopez M
Citation(s) 22851695
Submission date Feb 03, 2012
Last update date May 15, 2019
Contact name Claes M Gustafsson
Organization name Gothenburg University
Department Department of Medical biochemistry and cell biology
Street address Medcinaregatan 9A
City Gothenburg
ZIP/Postal code 405 30
Country Sweden
Platforms (1)
GPL13988 Illumina HiSeq 2000 (Schizosaccharomyces pombe)
Samples (3)
GSM870204 non antibody control ChIPseq
GSM870205 RNA Polymerase II CTD domain ChIPseq
GSM870206 Mediator subunit Med7 ChIPseq
This SubSeries is part of SuperSeries:
GSE35527 Mediator promotes CENP-A incorporation at fission yeast centromeres
SRA SRP010743
BioProject PRJNA156087

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Supplementary file Size Download File type/resource
GSE35524_RAW.tar 110.7 Mb (http)(custom) TAR (of BED, WIG)
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Raw data are available in SRA
Processed data provided as supplementary file

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