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Series GSE66830 Query DataSets for GSE66830
Status Public on May 06, 2016
Title The interaction of PRC2 with RNA or chromatin is mutually antagonistic [ChIP-Seq]
Organism Mus musculus
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary Polycomb repressive complex 2 (PRC2) maintains developmental regulator genes in a repressed state through methylation of histone H3 at lysine 27 (H3K27me3) and is necessary for cell differentiation. We and others have previously found that the PRC2 subunit Suz12 interacts with RNA in vitro and other studies have shown that Ezh2 and Jarid2 also possess RNA binding function. The interaction of PRC2 with RNA has been suggested to regulate PRC2 targeting or enzymatic activity, but the RNAs directly bound by PRC2 in cells, and the role of each PRC2 RNA binding subunit, remain unclear. We have used different CLIP techniques, which use UV-crosslinking to allow detection of direct Suz12-RNA interactions as they occur in living mouse ES cells. Suz12 binds nascent RNA and has a preference for interaction with the 3’UTR, showing it does have binding specificity in cells. RNAs bound by Suz12 at the 3’UTR encode developmental regulator genes. Suz12 remains bound to RNA upon deletion of Ezh2 or Jarid2 showing that it binds RNA independently of other PRC2 subunits. We also show that binding of Suz12 to RNA or chromatin is mutually inhibitory. Although Ezh2 and Jarid2 also bind RNA, Ezh2 and Jarid2 deletion causes an increase in Suz12 RNA binding, without changing its specificity, which reflects the loss of Suz12 from chromatin. Similarly, disruption of Suz12-RNA interactions by RNA polymerase II inhibition or RNase treatment increases Suz12 binding to chromatin. These results therefore suggest that Suz12 acts as an RNA sensor, binding to the 3’UTR of nascent RNAs and modulating the interaction of PRC2 with chromatin.
Overall design Suz12, and Ezh2 ChIP-seq experiments before and after Tamoxifen treatment of Mus musculus Ezh2 fl/fl Stem Cells.
Contributor(s) Beltran M, Yates CM, Viiri K, Skalska L, Reis F, Fisher CL, Dawson M, Sibley CR, Ule J, Jenner RG
Citation(s) 27197219
Submission date Mar 12, 2015
Last update date May 15, 2019
Contact name Richard Jenner
Organization name UCL Cancer Institute
Department Cancer Biology
Lab Regulatory Genomics
Street address 72 Huntley Street
City London
ZIP/Postal code WC1E 6BT
Country United Kingdom
Platforms (1)
GPL17021 Illumina HiSeq 2500 (Mus musculus)
Samples (12)
GSM1632842 Suz12_minusTmx
GSM1632843 Ezh2_minusTmx
GSM1632844 input_minusTmx
This SubSeries is part of SuperSeries:
GSE66831 The interaction of PRC2 with RNA or chromatin is mutually antagonistic
BioProject PRJNA278059
SRA SRP056117

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Supplementary file Size Download File type/resource
GSE66830_RAW.tar 9.1 Gb (http)(custom) TAR (of BW)
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Processed data provided as supplementary file
Raw data are available in SRA

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