Abstract

Phosphorylation of Ser/Thr residues is a well-established modulating mechanism of the pro-apoptotic function of the BH3-only protein Bim. However, nothing is known about the putative tyrosine phosphorylation of this Bcl-2 family member and its potential impact on Bim function and subsequent Bax/Bak-mediated cytochrome c release and apoptosis. As we have previously shown that the tyrosine kinase Lyn could behave as an anti-apoptotic molecule, we investigated whether this Src family member could directly regulate the pro-apoptotic function of Bim. In the present study, we show that Bim is phosphorylated onto tyrosine residues 92 and 161 by Lyn, which results in an inhibition of its pro-apoptotic function. Mechanistically, we show that Lyn-dependent tyrosine phosphorylation of Bim increases its interaction with anti-apoptotic members such as Bcl-xL, therefore limiting mitochondrial outer membrane permeabilization and subsequent apoptosis. Collectively, our data uncover one molecular mechanism through which the oncogenic tyrosine kinase Lyn negatively regulates the mitochondrial apoptotic pathway, which may contribute to the transformation and/or the chemotherapeutic resistance of cancer cells.

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Acknowledgements

This work was supported by grants from the Fondation ARC pour la Recherche sur le Cancer (#PJA 20131200392 and #PGA1RF201702053889), the Fondation pour la Recherche Médicale (DMP20101120387), the INSERM, the Ministere de la Recherche, and the University of Nice-Sophia-Antipolis. This work and LEA were funded by the French Government (National Research Agency, ANR) through the “Investments for the Future” LABEX SIGNALIFE: program reference #ANR-11-LABX-0028-01. We thank Frederic Larbret for technical support for FACS sorting. We thank Dr. Simon Cook for GST-Bim constructs, Dr. Andreas Villunger for kindly providing the WEHI-231 cell line, Dr. David Rubinsztein for the plasmid encoding human BimEL, and Drs. Doug Green and Stephen Tait for HeLa cells stably expressing Smac-GFP. We thank Drs. Eric Eldering and Fabio Brocco for insights and helpful discussion. We thank the Conseil général des AM et de la région PACA et Corse for its financial support.

Author contributions

SM and PA conceived the project. LEA, EV, PG, and SM conducted the experiments and were assisted by PC, LS, SO, EP. FG and PPJ contributed to the BRET experiments. SM wrote the paper. PA, J-ER, LEA, BB-M, AJ, GR, FL, FG, and PPJ reviewed and edited the paper. SM and PA supervised the study and obtained funding.

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Author notes

  1. Patrick Auberger and Sandrine Marchetti contributed equally to this work.

Affiliations

  1. Université Côte d’Azur, INSERM, C3M, Nice, France

    • Lazaro E. Aira
    • , Elodie Villa
    • , Pascal Colosetti
    • , Parvati Gamas
    • , Laurie Signetti
    • , Sandrine Obba
    • , Emma Proics
    • , Béatrice Bailly-Maitre
    • , Arnaud Jacquel
    • , Guillaume Robert
    • , Frédéric Luciano
    • , Jean-Ehrland Ricci
    • , Patrick Auberger
    •  & Sandrine Marchetti
  2. CRCINA, UMR 1232 INSERM, Université de Nantes, Université d’Angers, Institut de Recherche en Santé-Université de Nantes, 8 Quai Moncousu - BP 70721, 44007, Nantes Cedex 1, France

    • Fabien Gautier
    •  & Philippe P. Juin
  3. Institut de Cancérologie de l’Ouest, Bvd J Monod, Site René Gauducheau, 44805, Saint-Herblain, France

    • Fabien Gautier
    •  & Philippe P. Juin

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Correspondence to Sandrine Marchetti.

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DOI

https://doi.org/10.1038/s41388-017-0112-0