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Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS
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2017 (English)In: eLIFE, E-ISSN 2050-084X, Vol. 6, article id e27574Article in journal (Refereed) Published
Abstract [en]

The temporal regulation of protein abundance and post-translational modifications is a key feature of cell division. Recently, we analysed gene expression and protein abundance changes during interphase under minimally perturbed conditions (Ly et al., 2014, 2015). Here, we show that by using specific intracellular immunolabelling protocols, FACS separation of interphase and mitotic cells, including mitotic subphases, can be combined with proteomic analysis by mass spectrometry. Using this PRIMMUS (PRoteomic analysis of Intracellular iMMUnolabelled cell Subsets) approach, we now compare protein abundance and phosphorylation changes in interphase and mitotic fractions from asynchronously growing human cells. We identify a set of 115 phosphorylation sites increased during G2, termed ‘early risers’. This set includes phosphorylation of S738 on TPX2, which we show is important for TPX2 function and mitotic progression. Further, we use PRIMMUS to provide the first a proteome-wide analysis of protein abundance remodeling between prophase, prometaphase and anaphase.

Place, publisher, year, edition, pages
eLife Sciences Publications Ltd , 2017. Vol. 6, article id e27574
Keyword [en]
membrane protein, rrm2 protein, tpx2 protein, unclassified drug, analytic method, Article, cell cycle progression, controlled study, flow cytometry, fluorescence activated cell sorting, gene overexpression, high performance liquid chromatography, human, human cell, immunofluorescence microscopy, immunofluorescence test, interphase, liquid chromatography-mass spectrometry, mass spectrometry, mitosis, protein expression, protein phosphorylation, PRoteomic analysis of Intracellular iMMUnolabelled cell Subsets, proteomics, Western blotting
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-227081DOI: 10.7554/eLife.27574Scopus ID: 2-s2.0-85032944461OAI: oai:DiVA.org:kth-227081DiVA, id: diva2:1206645
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QC 20180517

Available from: 2018-05-17 Created: 2018-05-17 Last updated: 2018-05-17Bibliographically approved

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Lundberg, Emma

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