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Series GSE59518 Query DataSets for GSE59518
Status Public on Sep 16, 2015
Title The Mediator complex subunit MED23 couples H2B mono-ubiquitination to transcriptional control and cell fate determination
Organism Mus musculus
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The Mediator complex orchestrates multiple transcription factors with the Pol II apparatus for precise transcriptional control. However, its interplay with the surrounding chromatin remains poorly understood. Here, we analyze differential histone modifications between WT and MED23-/- (KO) cells and identify H2B mono-ubiquitination at lysine 120 (H2Bub) as a MED23-dependent histone modification. Using tandem affinity purification and mass spectrometry, we find that MED23 associates with the RNF20/40 complex, the enzyme for H2Bub, and show that this association is critical for the recruitment of RNF20/40 to chromatin. In a cell-free system, Mediator directly and substantially increases H2Bub on recombinant chromatin through its cooperation with RNF20/40 and the PAF complex. Integrative genome-wide analyses show that MED23 depletion specifically reduces H2Bub on a subset of MED23-contolled genes. Importantly, MED23-coupled H2Bub levels are oppositely regulated during myogenesis and lung carcinogenesis. In sum, these results establish a mechanistic link between the Mediator complex and a critical chromatin modification in coordinating transcription with cell growth and differentiation.
Overall design To examine the enrichment of H2B ubiquitination, Pol II, H3K4me3, H3K79me3 in WT and KO MED23 MEF cells, we performed H2Bub ChIP-seq, Pol II ChIP-seq, H3K4me3 ChIP-seq and H3K79me3 ChIP-seq assays. 10 high-throughput sequencing data were deposited and WT, KO input data were controls for peak calling.
Contributor(s) Yao X, Fu X, Wang G
Citation(s) 26330467
Submission date Jul 17, 2014
Last update date May 15, 2019
Contact name Gang Wang
Organization name Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences
Department Institute of Biochemistry and Cell Biology, State Key Laboratory of Cell Biology,
Lab Gang Wang's Lab
Street address Yueyang Road 320
City Shanghai
ZIP/Postal code 200031
Country China
Platforms (2)
GPL11002 Illumina Genome Analyzer IIx (Mus musculus)
GPL13112 Illumina HiSeq 2000 (Mus musculus)
Samples (10)
GSM1438697 pol2_WT_ChIPSeq
GSM1438698 pol2_KO_ChIPSeq
GSM1438699 h2bub_WT_ChIPSeq
BioProject PRJNA255494
SRA SRP044423

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Supplementary file Size Download File type/resource
GSE59518_RAW.tar 2.6 Gb (http)(custom) TAR (of WIG)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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