A FRET biosensor reveals spatiotemporal activation and functions of aurora kinase A in living cells.
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.
CNRS, UMR 6290, Rennes 35043, France.|Université de Rennes 1, Institut de Génétique et Développement de Rennes, Rennes 35043, France.
Giulia
G
Bertolin
CNRS, UMR 6290, Rennes 35043, France.|Université de Rennes 1, Institut de Génétique et Développement de Rennes, Rennes 35043, France.
Florian
F
Sizaire
CNRS, UMR 6290, Rennes 35043, France.|Université de Rennes 1, Institut de Génétique et Développement de Rennes, Rennes 35043, France.
Gaëtan
G
Herbomel
CNRS, UMR 6290, Rennes 35043, France.|Université de Rennes 1, Institut de Génétique et Développement de Rennes, Rennes 35043, France.|Equipe labéllisée Ligue Contre Le Cancer 2014-2016, Rennes 35043, France.
David
D
Reboutier
CNRS, UMR 6290, Rennes 35043, France.|Université de Rennes 1, Institut de Génétique et Développement de Rennes, Rennes 35043, France.|Equipe labéllisée Ligue Contre Le Cancer 2014-2016, Rennes 35043, France.
Claude
C
Prigent
CNRS, UMR 6290, Rennes 35043, France.|Université de Rennes 1, Institut de Génétique et Développement de Rennes, Rennes 35043, France.|Microscopy Rennes Imaging Centre, Biosit, Université de Rennes 1, Rennes 35043, France.
Marc
M
Tramier
D064096:Aurora Kinase A / Q000378:metabolism*; D015374:Biosensing Techniques; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004789:Enzyme Activation; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism
D018797:Cell Cycle Proteins; C526596:Cep192 protein, human; D002868:Chromosomal Proteins, Non-Histone; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; C122167:TPX2 protein, human; C576917:AURKA protein, human; D064096:Aurora Kinase A