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CDK8 and MED12 knockdown in R1 mouse ES cells

ABSTRACT: Transcriptional profiling of R1 mouse ES cells infected with non-targeting control (NTC) shRNA and two different shRNA sequences against Cdk8 (shCdk8-1 and shCdk-2) and Med12 (shMed12-1 and shMed12-2).

ORGANISM(S): Mus musculus

PROVIDER: GSE30816 | GEO | 2012/03/05

SECONDARY ACCESSION(S): PRJNA154571

REPOSITORIES: GEO

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CDK8 and MED12 knockdown in R1 mouse ES cells

Project description:Transcriptional profiling of R1 mouse ES cells infected with non-targeting control (NTC) shRNA and two different shRNA sequences against Cdk8 (shCdk8-1 and shCdk-2) and Med12 (shMed12-1 and shMed12-2). Multi-condition experiment comparing shNTC versus shCdk8-1 and shCdk8-2 after 8 days (3 biological replicates each) and shNTC versus shCdk8-1, shCdk8-2, shMed12-1, and shMed12-2 after 13 days (3 biological replicates each); in total 24 samples profiled.

2012-03-05 | E-GEOD-30816 | biostudies-arrayexpress

CDK8 knockdown in HT-29 cells and CDK8 & MED12 knockdown in R1 cells

Project description:This SuperSeries is composed of the following subset Series: GSE30815: CDK8 knockdown in HT-29 human colon cancer cells GSE30816: CDK8 and MED12 knockdown in R1 mouse ES cells Refer to individual Series

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A precisely positioned MED12 activation helix stimulates CDK8 kinase activity

Project description:The Mediator kinase module regulates eukaryotic transcription by phosphorylating transcription-related targets and by modulating the association of Mediator and RNA Polymerase II. The activity of its catalytic core, cyclin-dependent kinase 8 (CDK8), is controlled by Cyclin C and MED12, with its deregulation contributing to numerous malignancies. Here, we combine in vitro biochemistry, crosslinking coupled to mass spectrometry and in vivo studies to describe the binding location of the N-terminal segment of MED12 on the CDK8/Cyclin C complex and to gain mechanistic insights into the activation of CDK8 by MED12. Our data demonstrate that the N-terminal portion of MED12 wraps around CDK8, whereby it positions an "activation helix" close to the T-loop of CDK8 for its activation. Intriguingly, mutations in the activation helix that are frequently found in cancers do not diminish the affinity of MED12 for CDK8, yet likely alter the exact positioning of the activation helix. Furthermore, we find the transcriptome-wide gene expression changes in human cells that result from a mutation in the MED12 activation helix to correlate with deregulated genes in breast and colon cancer. Last, functional assays in presence of kinase inhibitors reveal that binding of MED12 remodels the active site of CDK8 and thereby precludes the inhibition of ternary CDK8 complexes by type-II kinase inhibitors. Taken together, our results not only allow us to propose a revised model of how CDK8 activity is regulated by MED12, but they also offer a path forward in developing small molecules that target CDK8 in its MED12-bound form.

2020-01-04 | GSE135458 | GEO

Cdk8, Cyclin C, Med12 or Med13 depletion effect on gene expression in Drosophila S2 cells

Project description:Expression profiling following depletion of Mediator Cdk8 module subunits Cdk8, Cyclin C (CycC), Med12 and Med13 72 hours after dsRNA treatment of Drosophila melanogaster S2 cells. Results provide insight into the role of individual Cdk8 module subunits in regulation of transcription. 22 samples. 2 Cdk8 dsRNA, 4 CycC dsRNA, 4 Med12 dsRNA, 4 Med13 dsRNA, 8 control samples including 4 Luciferase (Luc) dsRNA and 4 GFP dsRNA

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A precisely positioned MED12 activation helix stimulates CDK8 kinase activity

Project description:Chemical cross-linking coupled to mass spectrometry was used to study binary and ternary complexes involving cyclin-dependent kinase 8 (CDK8), cyclin-C, and subunit 12 of the Mediator complex (MED12). Cross-linking was performed using different cross-linking chemistries: (1) disuccinimidyl suberate (DSS); (2) a combination of pimelic acid dihydrazide (PDH) and the coupling reagent, DMTMM.

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Cdk8, Cyclin C, Med12 or Med13 depletion effect on gene expression in Drosophila S2 cells

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Mouse R1 ES cell SAGE library

Project description:Mouse R1 ES cell SAGE library Keywords: other

2005-09-01 | GSE3233 | GEO

Embryoid Bodies from Mouse R1 ES - 9 day

Project description:Embryoid Bodies from Mouse R1 ES - 9 day Keywords: other

2005-07-28 | GSE3007 | GEO

DR4 Control for J1, R1 and V6.5 ES

Project description:This sample is a control for the J1, R1 and V6.5 ES samples. The DR4 cell type acts as a feeder cell for stem cells and as such is a small contaminant of the embryonic stem cell growth environment. Keywords: other

2005-09-01 | GSE3232 | GEO

Mus musculus

Project description:CDK8 and MED12 knockdown in R1 mouse ES cells

| PRJNA154571 | ENA