Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of human male embryonic stem cell line line grown in standard conditions, with feeder cells and treated with the signalling sphingolipid sphingosine-1-phosphate reveals S1P mediates key targets associated with survival, proliferation and pluripotency

ABSTRACT: Human embryonic stem cells (hESCs) replicate by the process of self-renewal, whilst maintaining their pluripotency. Understanding the pathways involved in the regulation of this self-renewal process will assist in developing fully-defined conditions for the proliferation of hESCS required for therapeutic applications. We previously demonstrated a role for Sphingosine-1-phosphate (S1P) in the survival and proliferation of hESCs. The present study investigates further key signalling pathways and the downstream targets of S1P. Microarrays were used to examine changes in gene expression invoked by treatment of human embryonic stem cells grown upon different matrices Experiment Overall Design: Human embryonic stem cells (Sheff 4) were grown upon MEFs. One group were treated with S1P. Each group consisted of three replicates which were hybridised to Human U133 plus 2 Affymetrix GeneChips

ORGANISM(S): Homo sapiens

SUBMITTER: Paul Heath

PROVIDER: E-GEOD-7896 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Sphingosine-1-phosphate mediates transcriptional regulation of key targets associated with survival, proliferation, and pluripotency in human embryonic stem cells.

Avery Katie K Avery Stuart S Shepherd Jill J Heath Paul R PR Moore Harry H

Stem cells and development 20081201 6

Human embryonic stem cells (hESCs) replicate in vitro by the process of self-renewal, whilst maintaining their pluripotency. Understanding the pathways involved in the regulation of this process will assist in developing fully-defined conditions for the robust proliferation of hESCs necessary for therapeutic applications. We previously demonstrated that sphingosine-1-phosphate (S1P) plays an important role in survival and proliferation of hESCs. and here the key signaling pathways and downstream ...[more]

PMID: 18393631

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Project description:Here, we performed RNA-interactome capture (RIC) on nuclear fractions from human embryonic stem cells (hESCs). The poly(A)+ RNA-bound proteome was determined by UV light-mediated cross-linking (CL) of RNAs to proteins in living cells, followed by nuclei isolation, oligo(dT) purification of poly(A)-RNA-protein complexes, and mass spectrometry analysis of captured proteins. As a control, we applied a similar strategy to non-cross-linked (non-CL) samples. RIC was performed in four independent biological replicates. This data accompanies the manuscript: "Uncovering the RNA-binding protein landscape in the pluripotency network of human embryonic stem cells". Abstract: "Embryonic stem cell (ESC) self-renewal and cell-fate decisions are driven by a broad array of molecular signals. While transcriptional regulators have been extensively studied in human ESCs (hESCs), the extent to which RNA-binding proteins (RBPs) contribute to human pluripotency remains unclear. Here, we carry out a proteome-wide screen and identify 810 proteins that directly bind RNA in hESCs. We reveal that RBPs are preferentially expressed in hESCs and dynamically regulated during exit from pluripotency and early lineage specification. Moreover, we show that nearly 200 RBPs are affected by knockdown of OCT4, a master regulator of pluripotency, several dozen of which are directly bound by this factor. Intriguingly, over 20 percent of the proteins detected in our study are putative DNA- and RNA-binding proteins (DRBPs), among them key transcription factors (TFs). Using fluorescently labeled RNA and seCLIP (single-end enhanced crosslinking and immunoprecipitation) experiments, we discover that the pluripotency-associated STAT3 and OCT4 TFs interact with RNA in hESCs and confirm the direct binding of STAT3 to the conserved NORAD long-noncoding RNA. Taken together, our findings indicate that RBPs have a more widespread role in human pluripotency than previously appreciated, reinforcing the importance of post-transcriptional regulation in stem cell biology".

Dataset Information

Transcription profiling of human male embryonic stem cell line line grown in standard conditions, with feeder cells and treated with the signalling sphingolipid sphingosine-1-phosphate reveals S1P mediates key targets associated with survival, proliferation and pluripotency

Publications

Sphingosine-1-phosphate mediates transcriptional regulation of key targets associated with survival, proliferation, and pluripotency in human embryonic stem cells.

Similar Datasets

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