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| Status |
Public on Jun 16, 2021 |
| Title |
The Cdk8 kinase module regulates Mediator-RNA polymerase II interaction |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
The Cdk8 kinase module (CKM) is a dissociable part of the coactivator complex Mediator that regulates RNA polymerase II (Pol II transcription. The CKM has negative and positive functions in gene transcription that remain poorly understood at the mechanistic level. Here, we prepare recombinant CKM from the yeast Saccharomyces cerevisiae and show that it binds core Mediator (cMed) to sterically inhibit cMed binding to the Pol II preinitiation complex (PIC) in vitro. We further show that the Cdk8 kinase activity of CKM counteracts CKM-cMed interaction,thereby releasing CKM and enabling Mediator to bind the PIC. Finally, we report that the kinase activity of Cdk8 is required for gene activation during heat shock in vivo, but not under steady state growth conditions. These results converge with previous literature on a model for CKM function. In this model, CKM negatively regulates Mediator function at upstream activating sequences by preventing Mediator binding to the PIC at the promoter. During gene activation, Cdk8 kinase activity may release Mediator and allow its binding to the PIC, thereby stimulating transcription initiation and accounting for the positive function of CKM.
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| |
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| Overall design |
4tU-seq in yeast (Saccharomyces cerevisiae) Srb10 analogue-sensitive cells (Srb10 as-1) upon Srb10 inhibition
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| |
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| Contributor(s) |
Osman S, Maier K, Mohammad E, Lidschreiber M, Cramer P |
| Citation(s) |
33933450 |
| |
| Submission date |
Nov 09, 2020 |
| Last update date |
Jun 16, 2021 |
| Contact name |
Eusra Mohammad |
| E-mail(s) |
[email protected]
|
| Organization name |
Max Planck Institute for Biophysical Chemistry
|
| Street address |
Am Faßberg 11
|
| City |
Göttingen |
| ZIP/Postal code |
37077 |
| Country |
Germany |
| |
|
| Platforms (1) |
| GPL19756 |
Illumina NextSeq 500 (Saccharomyces cerevisiae) |
|
| Samples (8)
|
|
| Relations |
| BioProject |
PRJNA675650 |
| SRA |
SRP291785 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE161140_SteadyStatevsHeatShock.DMSO.res.table.DE.txt.gz |
275.7 Kb |
(ftp)(http) |
TXT |
| GSE161140_SteadyStatevsHeatShock.NAPP1.res.table.DE.txt.gz |
267.8 Kb |
(ftp)(http) |
TXT |
| GSE161140_cerevisiae.feature.anno.orf.txt.gz |
110.0 Kb |
(ftp)(http) |
TXT |
| GSE161140_cerevisiae.feature.orfs.normalised.htseq.counts.HeatShock.txt.gz |
138.4 Kb |
(ftp)(http) |
TXT |
| GSE161140_cerevisiae.feature.orfs.normalised.htseq.counts.SteadyState.txt.gz |
147.1 Kb |
(ftp)(http) |
TXT |
| GSE161140_cerevisiae.feature.orfs.normalised.htseq.counts.SteadyStateVsHeatShock.1-NA-PP1.txt.gz |
142.7 Kb |
(ftp)(http) |
TXT |
| GSE161140_cerevisiae.feature.orfs.normalised.htseq.counts.SteadyStateVsHeatShock.DMSO.txt.gz |
141.8 Kb |
(ftp)(http) |
TXT |
| GSE161140_cerevisiae.feature.orfs.raw.htseq.counts.txt.gz |
99.4 Kb |
(ftp)(http) |
TXT |
| GSE161140_heat.shock.response.genes.id.txt.gz |
139 b |
(ftp)(http) |
TXT |
| GSE161140_heat.shock.response.genes.txt.gz |
173 b |
(ftp)(http) |
TXT |
| GSE161140_induced.heat.shock.response.genes.id.txt.gz |
131 b |
(ftp)(http) |
TXT |
| GSE161140_induced.heat.shock.response.genes.txt.gz |
141 b |
(ftp)(http) |
TXT |
| GSE161140_normalizationFactors.DESeq.HeatShock.txt.gz |
160 b |
(ftp)(http) |
TXT |
| GSE161140_normalizationFactors.DESeq.SteadyState.txt.gz |
163 b |
(ftp)(http) |
TXT |
| GSE161140_normalizationFactors.DESeq.SteadyStateVsHeatShock.1-NA-PP1.txt.gz |
188 b |
(ftp)(http) |
TXT |
| GSE161140_normalizationFactors.DESeq.SteadyStateVsHeatShock.DMSO.txt.gz |
181 b |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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