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| Status |
Public on Mar 21, 2016 |
| Title |
The cohesin associated protein Wapal is required for proper polycomb-mediated gene silencing [Ring1b ChIP-seq] |
| Organism |
Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
The cohesin complex consists of multiple core subunits that play critical roles in mitosis and transcriptional regulation. The cohesin-associated protein Wapal plays a central role in offloading cohesin to facilitate sister chromatid separation, but its role in regulating mammalian gene expression is not understood. We used embryonic stem cells (ESCs) as a model given the well-defined transcriptional regulatory circuits established through master transcription factors and epigenetic pathways that regulate their ability to maintain a pluripotent state. RNAi-mediated depletion of Wapal causes a loss of pluripotency, phenocopying loss of core cohesin subunits. Using chromatin immunoprecipitation coupled with next-generation sequencing (ChIP-seq) we determine that Wapal occupies genomic sites distal to genes in combination with CTCF and core cohesin subunits such as Rad21. Interestingly, genomic sites occupied by Wapal appear enriched for cohesin, implying Wapal does not offload cohesin at regions it occupies. Wapal depletion induces derepression of Polycomb Group (PcG) target genes without altering total levels of Polycomb-mediated histone modifications, implying that PcG enzymatic activity is preserved. By integrating ChIP-seq and gene expression changes data we identify that Wapal binding is enriched at the promoters of PcG silenced genes and is required for proper Polycomb Repressive Complex 2 (PRC2) recruitment. Lastly, we demonstrate that Wapal is required for the interaction of a distal cis-regulatory element (CRE) with the c-Fos promoter. Collectively, this work indicates that Wapal plays a critical role in silencing of PcG target genes through the interaction of distal CREs with promoters.
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| |
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| Overall design |
ChIP-seq for Ring1b, before or after depletion of Wapal by RNAi
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| |
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| Contributor(s) |
Rao S |
| Citation(s) |
27087855 |
| Submission date |
Mar 02, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Sridhar Rao |
| E-mail(s) |
sridhar.rao@bcw.edu
|
| Phone |
4149373841
|
| Organization name |
BloodCenter of Wisconsin
|
| Department |
Blood Research Institute
|
| Street address |
8727 Watertown Plank Road
|
| City |
Milwaukee |
| State/province |
WI |
| ZIP/Postal code |
53226 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
|
| Samples (20)
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| This SubSeries is part of SuperSeries: |
| GSE63325 |
The cohesin associated factor Wapal is required for proper polycomb-mediated gene silencing |
|
| Relations |
| BioProject |
PRJNA314075 |
| SRA |
SRP071063 |
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