Quantitative proteomics analysis demonstrates post-transcriptional regulation of embryonic stem cell differentiation to hematopoiesis.

2.50
Hdl Handle:
http://hdl.handle.net/10541/68796
Title:
Quantitative proteomics analysis demonstrates post-transcriptional regulation of embryonic stem cell differentiation to hematopoiesis.
Authors:
Williamson, Andrew J K; Smith, Duncan L; Blinco, David; Unwin, Richard D; Pearson, Stella; Wilson, Claire L; Miller, Crispin J; Lancashire, Lee J; Lacaud, Georges; Kouskoff, Valerie; Whetton, Anthony D
Abstract:
Embryonic stem (ES) cells can differentiate in vitro to produce the endothelial and hematopoietic precursor, the hemangioblasts, which are derived from the mesoderm germ layer. Differentiation of Bry(GFP/+) ES cell to hemangioblasts can be followed by the expression of the Bry(GFP/+) and Flk1 genes. Proteomic and transcriptomic changes during this differentiation process were analyzed to identify mechanisms for phenotypic change during early differentiation. Three populations of differentiating Bry(GFP) ES cells were obtained by flow cytometric sorting, GFP-Flk1- (epiblast), GFP+Flk1- (mesoderm), and GFP+Flk1+ (hemangioblast). Microarray analyses and relative quantification two-dimensional LCLC-MS/MS on nuclear extracts were performed. We identified and quantified 2389 proteins, 1057 of which were associated to their microarray probe set. These included a variety of low abundance transcription factors, e.g. UTF1, Sox2, Oct4, and E2F4, demonstrating a high level of proteomic penetrance. When paired comparisons of changes in the mRNA and protein expression levels were performed low levels of correlation were found. A strong correlation between isobaric tag-derived relative quantification and Western blot analysis was found for a number of nuclear proteins. Pathway and ontology analysis identified proteins known to be involved in the regulation of stem cell differentiation, and proteins with no described function in early ES cell development were also shown to change markedly at the proteome level only. ES cell development is regulated at the mRNA and protein level.
Affiliation:
Stem Cell and Leukemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Kinnaird House, Kinnaird Road, Manchester M20 4QL, United Kingdom.
Citation:
Quantitative proteomics analysis demonstrates post-transcriptional regulation of embryonic stem cell differentiation to hematopoiesis. 2008, 7 (3):459-72 Mol. Cell Proteomics
Journal:
Molecular & Cellular Proteomics
Issue Date:
Mar-2008
URI:
http://hdl.handle.net/10541/68796
DOI:
10.1074/mcp.M700370-MCP200
PubMed ID:
18045800
Type:
Article
Language:
en
ISSN:
1535-9484
Appears in Collections:
Applied Computational Biology and Bioinformatics; All Paterson Institute for Cancer Research; Clinical and Experimental Pharmacology Group; School of Cancer and Imaging Sciences; Stem Cell and Haematopoiesis; Stem Cell Biology; Molecular Biology Core Facility

Full metadata record

DC FieldValue Language
dc.contributor.authorWilliamson, Andrew J K-
dc.contributor.authorSmith, Duncan L-
dc.contributor.authorBlinco, David-
dc.contributor.authorUnwin, Richard D-
dc.contributor.authorPearson, Stella-
dc.contributor.authorWilson, Claire L-
dc.contributor.authorMiller, Crispin J-
dc.contributor.authorLancashire, Lee J-
dc.contributor.authorLacaud, Georges-
dc.contributor.authorKouskoff, Valerie-
dc.contributor.authorWhetton, Anthony D-
dc.date.accessioned2009-05-22T13:14:06Z-
dc.date.available2009-05-22T13:14:06Z-
dc.date.issued2008-03-
dc.identifier.citationQuantitative proteomics analysis demonstrates post-transcriptional regulation of embryonic stem cell differentiation to hematopoiesis. 2008, 7 (3):459-72 Mol. Cell Proteomicsen
dc.identifier.issn1535-9484-
dc.identifier.pmid18045800-
dc.identifier.doi10.1074/mcp.M700370-MCP200-
dc.identifier.urihttp://hdl.handle.net/10541/68796-
dc.description.abstractEmbryonic stem (ES) cells can differentiate in vitro to produce the endothelial and hematopoietic precursor, the hemangioblasts, which are derived from the mesoderm germ layer. Differentiation of Bry(GFP/+) ES cell to hemangioblasts can be followed by the expression of the Bry(GFP/+) and Flk1 genes. Proteomic and transcriptomic changes during this differentiation process were analyzed to identify mechanisms for phenotypic change during early differentiation. Three populations of differentiating Bry(GFP) ES cells were obtained by flow cytometric sorting, GFP-Flk1- (epiblast), GFP+Flk1- (mesoderm), and GFP+Flk1+ (hemangioblast). Microarray analyses and relative quantification two-dimensional LCLC-MS/MS on nuclear extracts were performed. We identified and quantified 2389 proteins, 1057 of which were associated to their microarray probe set. These included a variety of low abundance transcription factors, e.g. UTF1, Sox2, Oct4, and E2F4, demonstrating a high level of proteomic penetrance. When paired comparisons of changes in the mRNA and protein expression levels were performed low levels of correlation were found. A strong correlation between isobaric tag-derived relative quantification and Western blot analysis was found for a number of nuclear proteins. Pathway and ontology analysis identified proteins known to be involved in the regulation of stem cell differentiation, and proteins with no described function in early ES cell development were also shown to change markedly at the proteome level only. ES cell development is regulated at the mRNA and protein level.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshCell Differentiation-
dc.subject.meshEmbryonic Stem Cells-
dc.subject.meshFetal Proteins-
dc.subject.meshGene Expression Profiling-
dc.subject.meshGene Expression Regulation, Developmental-
dc.subject.meshGreen Fluorescent Proteins-
dc.subject.meshHematopoiesis-
dc.subject.meshMetabolic Networks and Pathways-
dc.subject.meshMice-
dc.subject.meshProtein Array Analysis-
dc.subject.meshProteomics-
dc.subject.meshRNA, Messenger-
dc.subject.meshReproducibility of Results-
dc.subject.meshT-Box Domain Proteins-
dc.subject.meshTandem Mass Spectrometry-
dc.subject.meshTranscription, Genetic-
dc.subject.meshVascular Endothelial Growth Factor Receptor-2-
dc.titleQuantitative proteomics analysis demonstrates post-transcriptional regulation of embryonic stem cell differentiation to hematopoiesis.en
dc.typeArticleen
dc.contributor.departmentStem Cell and Leukemia Proteomics Laboratory, Faculty of Medical and Human Sciences, University of Manchester, Kinnaird House, Kinnaird Road, Manchester M20 4QL, United Kingdom.en
dc.identifier.journalMolecular & Cellular Proteomicsen

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