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A FRET biosensor reveals spatiotemporal activation and functions of aurora kinase A in living cells

Author

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  • Giulia Bertolin

    (CNRS, UMR 6290
    Université de Rennes 1, Institut de Génétique et Développement de Rennes)

  • Florian Sizaire

    (CNRS, UMR 6290
    Université de Rennes 1, Institut de Génétique et Développement de Rennes)

  • Gaëtan Herbomel

    (CNRS, UMR 6290
    Université de Rennes 1, Institut de Génétique et Développement de Rennes)

  • David Reboutier

    (CNRS, UMR 6290
    Université de Rennes 1, Institut de Génétique et Développement de Rennes
    Equipe labéllisée Ligue Contre Le Cancer 2014–2016)

  • Claude Prigent

    (CNRS, UMR 6290
    Université de Rennes 1, Institut de Génétique et Développement de Rennes
    Equipe labéllisée Ligue Contre Le Cancer 2014–2016)

  • Marc Tramier

    (CNRS, UMR 6290
    Université de Rennes 1, Institut de Génétique et Développement de Rennes
    Microscopy Rennes Imaging Centre, Biosit, Université de Rennes 1)

Abstract

Overexpression of AURKA is a major hallmark of epithelial cancers. It encodes the multifunctional serine/threonine kinase aurora A, which is activated at metaphase and is required for cell cycle progression; assessing its activation in living cells is mandatory for next-generation drug design. We describe here a Förster’s resonance energy transfer (FRET) biosensor detecting the conformational changes of aurora kinase A induced by its autophosphorylation on Thr288. The biosensor functionally replaces the endogenous kinase in cells and allows the activation of the kinase to be followed throughout the cell cycle. Inhibiting the catalytic activity of the kinase prevents the conformational changes of the biosensor. Using this approach, we discover that aurora kinase A activates during G1 to regulate the stability of microtubules in cooperation with TPX2 and CEP192. These results demonstrate that the aurora kinase A biosensor is a powerful tool to identify new regulatory pathways controlling aurora kinase A activation.

Suggested Citation

  • Giulia Bertolin & Florian Sizaire & Gaëtan Herbomel & David Reboutier & Claude Prigent & Marc Tramier, 2016. "A FRET biosensor reveals spatiotemporal activation and functions of aurora kinase A in living cells," Nature Communications, Nature, vol. 7(1), pages 1-16, November.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms12674
    DOI: 10.1038/ncomms12674
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    Cited by:

    1. Hela Benaissa & Karim Ounoughi & Isabelle Aujard & Evelyne Fischer & Rosette Goïame & Julie Nguyen & Alison G. Tebo & Chenge Li & Thomas Saux & Giulia Bertolin & Marc Tramier & Lydia Danglot & Nicolas, 2021. "Engineering of a fluorescent chemogenetic reporter with tunable color for advanced live-cell imaging," Nature Communications, Nature, vol. 12(1), pages 1-17, December.

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