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| Status |
Public on Feb 26, 2016 |
| Title |
24hr CA treatment vs. DMSO in HCT116 cells (from 'Identification of CDK8 and CDK19 substrates in human cells using cortistatin A and quantitative phosphoproteomics') |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Cortistatin A (CA) is a highly selective inhibitor of the Mediator kinases CDK8 and CDK19. Using CA, we report here the first large-scale identification of Mediator kinase substrates in human cells (HCT116). Among over 16,000 quantified phosphosites, we identified 78 high-confidence Mediator kinase targets within 64 proteins, including DNA-binding transcription factors and proteins associated with chromatin, DNA repair, and RNA polymerase II. Although RNA-Seq data correlated with Mediator kinase targets, CA effects on gene expression were limited and distinct from CDK8 or CDK19 knockdown. Quantitative proteome analyses, which tracked about 7,000 proteins across six time points (0 – 24h), revealed that CA selectively affected pathways implicated in inflammation, growth, and metabolic regulation; contrary to expectations, increased turnover of Mediator kinase targets was not generally observed. Collectively, these data support Mediator kinases as regulators of chromatin and RNA polymerase II activity and suggest cellular roles beyond transcription, including metabolism and DNA repair.
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| Overall design |
HCT116 cells were treated with either 100nM CA or DMSO in biological triplicate for each population (6 samples total). Treatment was for 24h for compound and vehicle.
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| Contributor(s) |
Poss ZC, Taatjes DJ |
| Citation(s) |
27050516 |
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| Submission date |
Feb 25, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Dylan Taatjes |
| Organization name |
University of Colorado, Boulder
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| Department |
Molecular and Cell Biology
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| Lab |
Taatjes Lab
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| Street address |
3415 Colorado Ave.
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| City |
Boulder |
| State/province |
Colorado |
| ZIP/Postal code |
80303 |
| Country |
USA |
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| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA313158 |
| SRA |
SRP070815 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE78506_dmso_vs_ca_24Hour_geneExp.diff.gz |
868.9 Kb |
(ftp)(http) |
DIFF |
SRA Run Selector |
| Processed data are available on Series record |
| Raw data are available in SRA |
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