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| Status |
Public on Jan 12, 2017 |
| Title |
RNA binds to a CBP regulatory motif to stimulate histone acetylation and transcription |
| Organism |
Mus musculus |
| Experiment type |
Other
Non-coding RNA profiling by high throughput sequencing
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| Summary |
CBP/p300 are transcription co-activators whose binding is a signature of enhancers, cis-regulatory elements that control patterns of gene expression in multicellular organisms. Active enhancers produce bi-directional enhancer RNAs (eRNAs) and display CBP/p300 dependent histone acetylation. Here, we demonstrate that CBP binds directly to RNAs in vivo and in vitro. RNAs bound to CBP in vivo include a large number of eRNAs. Using steady-state histone acetyltransferase (HAT) assays we show that an RNA binding region in the HAT domain of CBP—a regulatory motif unique to CBP/p300—allows RNA to stimulate CBP’s HAT activity. At enhancers where CBP interacts with eRNAs, stimulation manifests in RNA-dependent changes in the histone acetylation mediated by CBP, such as H3K27ac, and by corresponding changes in gene expression. By interacting directly with CBP, eRNAs contribute to the unique chromatin structure at active enhancers, which in turn is required for regulation of target genes.
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| Overall design |
MEFs were treated with 4-thiouridine for 16h, irradiated with 365nm UV and CBP immunopreciptated. RNA species cross-linked to CBP were purified and sequenced. The series contains 2 biological replicates for CBP PAR-CLIP seq, and negative controls for background PAR-CLIP binding: nuclear flag-tagged GFP (nGFP) and the DNA binding domain of yeast protein Gal4p (Gal4-DBD) (two replicates each)
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| Contributor(s) |
Bose DA, Donahue G, Berger SL |
| Citation(s) |
28086087 |
| Submission date |
Dec 03, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Daniel Bose |
| Organization name |
University of Pennsylvania
|
| Department |
Cell and Developmental Biology
|
| Lab |
Berger Lab
|
| Street address |
3400 Civic Center Blvd., 9th floor
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (6)
|
| GSM1963690 |
nGFP background control (Replicate 2) |
| GSM1963691 |
Gal4-DBD background control (Replicate 1) |
| GSM1963692 |
Gal4-DBD background control (Replicate 2) |
|
| Relations |
| BioProject |
PRJNA305043 |
| SRA |
SRP066981 |
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