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Cell Cycle Oscillators pp 157–171Cite as

Spatiotemporal Investigation of Phosphorylation Events During Cell Cycle Progression

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1342))

Abstract

Polo-like kinase 1 (Plk1) is an essential kinase for mitotic commitment and progression through mitosis. In contrast to its well characterized roles during mitosis, the precise molecular events controlled by Plk1 during G2/M progression and their spatiotemporal regulation are still poorly elucidated. We recently investigated Plk1-dependent regulation of Cdc25C phosphatase, an activator of the master mitotic driver Cyclin B1-Cdk1. To this end, we generated a genetically encoded FRET (Förster Resonance Energy Transfer)-based Cdc25C phosphorylation biosensor to observe Cdc25 spatiotemporal phosphorylation during cell cycle progression in live single cell assays. Because this approach proved to be powerful, we provide here guidelines for the development of biosensors for any phosphorylation site of interest.

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Author information

Authors and Affiliations

  1. Sorbonne Universités, UPMC Paris VI, UFR927, 75005, Paris, France

    Lilia Gheghiani

  2. Institut Gustave Roussy, UMR 8200 CNRS, PR2, 114 rue Edouard Vaillant, 94805, Villejuif Cedex, France

    Olivier Gavet

Authors
  1. Lilia Gheghiani
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  2. Olivier Gavet
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Corresponding author

Correspondence to Olivier Gavet .

Editor information

Editors and Affiliations

  1. Department of Oncology, University of Oxford, Oxford, United Kingdom

    Amanda S. Coutts

  2. Scientific writer, London, United Kingdom

    Louise Weston

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Gheghiani, L., Gavet, O. (2016). Spatiotemporal Investigation of Phosphorylation Events During Cell Cycle Progression. In: Coutts, A., Weston, L. (eds) Cell Cycle Oscillators. Methods in Molecular Biology, vol 1342. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2957-3_8

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  • DOI: https://doi.org/10.1007/978-1-4939-2957-3_8

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2956-6

  • Online ISBN: 978-1-4939-2957-3

  • eBook Packages: Springer Protocols

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