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

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    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
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    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.
    Issue Date
    2008-03
    
    Metadata
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    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.
    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
    URI
    http://hdl.handle.net/10541/68796
    DOI
    10.1074/mcp.M700370-MCP200
    PubMed ID
    18045800
    Type
    Article
    Language
    en
    ISSN
    1535-9484
    ae974a485f413a2113503eed53cd6c53
    10.1074/mcp.M700370-MCP200
    Scopus Count
    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

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