Unknown,Transcriptomics,Genomics,Proteomics

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MEF wt versus Med23-/- KO after 20% Serum Stimulation

ABSTRACT: Affymetrix MOE430A arrays. Mouse embryonic fibroblasts (MEFs) from E11 passaged through crisis. Starved in 0.5% FBS for 16 h then stimulated by 20% serum final concentration for 30 minutes. The purpose was to identify genes affected by the loss of MED23 protein (subunit of the mediator complex). Samples were assayed in duplicate (Set 1 and Set 2) for starved state and serum state cells. Cells starved in low-serum (0.5%) for 16 h then add final conc. 20% FBS for 30 minutes. Harvest total RNA from duplicate samples. One sample set is kept starved and serves as the baseline. The other set is stimulated. This is done for both wild-type cell and the Med23-/- KO cells. We looked for gene expression fold changes greater than 2 (plus or minus) to find genes affected for serum stimulation by the loss of MED23, looking in detail at immediate early genes like Egr1.

ORGANISM(S): Mus musculus

SUBMITTER: Michael Balamotis 

PROVIDER: E-GEOD-15638 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Complexity in transcription control at the activation domain-mediator interface.

Balamotis Michael A MA   Pennella Mario A MA   Stevens Jennitte L JL   Wasylyk Bohdan B   Belmont Andrew S AS   Berk Arnold J AJ  

Science signaling 20090505 69

Transcript elongation by polymerase II paused at the Egr1 promoter is activated by mitogen-activated protein kinase phosphorylation of the ternary complex factor (TCF) ELK1 bound at multiple upstream sites and subsequent phospho-ELK1 interaction with mediator through the MED23 subunit. Consequently, Med23 knockout (KO) nearly eliminates Egr1 (early growth response factor 1) transcription in embryonic stem (ES) cells, leaving a paused polymerase at the promoter. Med23 KO did not, however, elimina  ...[more]

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