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    Quantitative phosphoproteome analysis of embryonic stem cell differentiation toward blood.

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    Authors
    Piazzi, M
    Williamson, Andrew J K
    Lee, Chia-Fang
    Pearson, Stella
    Lacaud, Georges
    Kouskoff, Valerie
    McCubrey, James A
    Cocco, L
    Whetton, Anthony D
    Affiliation
    Cell Signaling Laboratory, Department of Biomedical Science (DIBINEM), University of Bologna, Italy
    Issue Date
    2015-03-26
    
    Metadata
    Show full item record
    Abstract
    Murine embryonic stem (ES) cells can differentiate in vitro into three germ layers (endodermic, mesodermic, ectodermic). Studies on the differentiation of these cells to specific early differentiation stages has been aided by an ES cell line carrying the Green Fluorescent Protein (GFP) targeted to the Brachyury (Bry) locus which marks mesoderm commitment. Furthermore, expression of the Vascular Endothelial Growth Factor receptor 2 (Flk1) along with Bry defines hemangioblast commitment. Isobaric-tag for relative and absolute quantification (iTRAQTM) and phosphopeptide enrichment coupled to liquid chromatography separation and mass spectrometry allow the study of phosphorylation changes occurring at different stages of ES cell development using Bry and Flk1 expression respectively. We identified and relatively quantified 37 phosphoentities which are modulated during mesoderm-induced ES cells differentiation, comparing epiblast-like, early mesoderm and hemangioblast-enriched cells. Among the proteins differentially phosphorylated toward mesoderm differentiation were: the epigenetic regulator Dnmt3b, the protein kinase GSK3b, the chromatin remodeling factor Smarcc1, the transcription factor Utf1; as well as protein specifically related to stem cell differentiation, as Eomes, Hmga2, Ints1 and Rif1. As most key factors regulating early hematopoietic development have also been implicated in various types of leukemia, understanding the post-translational modifications driving their regulation during normal development could result in a better comprehension of their roles during abnormal hematopoiesis in leukemia.
    Citation
    Quantitative phosphoproteome analysis of embryonic stem cell differentiation toward blood. 2015: Oncotarget
    Journal
    Oncotarget
    URI
    http://hdl.handle.net/10541/555805
    PubMed ID
    25890499
    Type
    Article
    Language
    en
    ISSN
    1949-2553
    Collections
    All Paterson Institute for Cancer Research

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