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E-GEOD-49305 - Polycomb protein EED is required for selective silencing of pluripotency genes upon ESC differentiation

Status
Released on 28 August 2014, last updated on 1 September 2014
Organism
Mus musculus
Samples (24)
Array (1)
Protocols (6)
Description
Eed (embryonic ectoderm development) is a core component of the Polycomb Repressive Complex 2 (PRC2) which catalyzes the methylation of histone H3 lysine 27 (H3K27). Trimethylated H3K27 (H3K27me3) can act as a signal for PRC1 recruitment in the process of gene silencing and chromatin condensation. Previous studies with Eed KO ESCs revealed a failure to down-regulate a limited list of pluripotency factors in differentiating ESCs. Our aim was to analyze the consequences of Eed KO for ESC differentiation. To this end we first analyzed ESC differentiation in the absence of Eed and employed in silico data to assess pluripotency gene expression and H3K27me3 patterns. We linked these data to expression analyses of wildtype and Eed KO ESCs. We observed that in wildtype ESCs a subset of pluripotency genes including Oct4, Nanog, Sox2 and Oct4 target genes progressively gain H3K27me3 during differentiation. These genes remain expressed in differentiating Eed KO ESCs. This suggests that the deregulation of a limited set of pluripotency factors impedes ESC differentiation. Global analyses of H3K27me3 and Oct4 ChIP-seq data indicate that in ESCs the binding of Oct4 to promoter regions is not a general predictor for PRC2-mediated silencing during differentiation. However, motif analyses suggest a binding of Oct4 together with Sox2 and Nanog at promoters of genes that are PRC2-dependently silenced during differentiation. In summary, our data further characterize Eed function in ESCs by showing that Eed/PRC2 is essential for the onset of ESC differentiation. RNAs obtained from undifferentiated (d0) wild type and Eed KO ESCs and from day 3 (d3) and day 7 (d7) respective Ebs were subjected to Affymetrix Mouse Gene 1.0 ST Array. 24 samples in total.
Experiment type
transcription profiling by array 
Contacts
Albrecht M Müller, Martin Zenke, Matthias Becker, Nadine Obier, Pierre Cauchy, Qiong Lin, Vroni Hornich
Citation
Polycomb Protein EED is Required for Silencing of Pluripotency Genes upon ESC Differentiation. Obier N, Lin Q, Cauchy P, Hornich V, Zenke M, Becker M, M�ller AM. , PMID:25134795
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-49305.idf.txt
Sample and data relationshipE-GEOD-49305.sdrf.txt
Processed data (1)E-GEOD-49305.processed.1.zip
Array designA-AFFY-130.adf.txt
Links