Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein

Tilman Brummer, Mark Larance, Maria Teresa Herrera Abreu, Ruth J. Lyons, Paul Timpson, Christoph H. Emmerich, Emmy D. G. Fleuren, Gillian M. Lehrbach, Daniel Schramek, Michael Guilhaus, David E. James, Roger J. Daly (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    43 Citations (Scopus)

    Abstract

    Grb2-associated binder (Gab)2 functions downstream of a variety of receptor and cytoplasmic tyrosine kinases as a docking platform for specific signal transducers and performs important functions in both normal physiology and oncogenesis. Gab2 signalling is promoted by its association with specific receptors through the adaptor Grb2. However, the molecular mechanisms that attenuate Gab2 signals have remained unclear. We now demonstrate that growth factor-induced phosphorylation of Gab2 on two residues, S210 and T391, leads to recruitment of 14-3-3 proteins. Together, these events mediate negative-feedback regulation, as Gab2(S210A/T391A) exhibits sustained receptor association and signalling and promotes cell proliferation and transformation. Importantly, introduction of constitutive 14-3-3-binding sites into Gab2 renders it refractory to receptor activation, demonstrating that site-selective binding of 14-3-3 proteins is sufficient to terminate Gab2 signalling. Furthermore, this is associated with reduced binding of Grb2. This leads to a model where signal attenuation occurs because 14-3-3 promotes dissociation of Gab2 from Grb2, and thereby uncouples Gab2 from the receptor complex. This represents a novel regulatory mechanism with implications for diverse tyrosine kinase signalling systems.
    Original languageEnglish
    Pages (from-to)2305-2316
    Number of pages12
    JournalEMBO Journal
    Volume27
    Issue number17
    DOIs
    Publication statusPublished - 3 Sep 2008

    Fingerprint

    14-3-3 Proteins
    Phosphorylation
    Binding Sites
    Receptor Protein-Tyrosine Kinases
    Transducers
    Protein-Tyrosine Kinases
    Intercellular Signaling Peptides and Proteins
    Carcinogenesis
    Proteins
    Cell Proliferation
    Association reactions
    Physiology
    Cell proliferation
    Cytoplasmic and Nuclear Receptors
    Refractory materials
    Binders
    Chemical activation
    Feedback

    Keywords

    • 14-3-3 Proteins
    • Recombinant Proteins
    • Humans
    • Amino Acid Sequence
    • Feedback, Physiological
    • Models, Biological
    • Protein Binding
    • Binding Sites
    • Mutagenesis, Site-Directed
    • Multiprotein Complexes
    • Phosphorylation
    • Adaptor Proteins, Signal Transducing
    • Molecular Sequence Data
    • Sequence Homology, Amino Acid
    • Signal Transduction
    • Cell Line

    Cite this

    Brummer, T., Larance, M., Herrera Abreu, M. T., Lyons, R. J., Timpson, P., Emmerich, C. H., ... Daly, R. J. (2008). Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein. EMBO Journal, 27(17), 2305-2316. https://doi.org/10.1038/emboj.2008.159
    Brummer, Tilman ; Larance, Mark ; Herrera Abreu, Maria Teresa ; Lyons, Ruth J. ; Timpson, Paul ; Emmerich, Christoph H. ; Fleuren, Emmy D. G. ; Lehrbach, Gillian M. ; Schramek, Daniel ; Guilhaus, Michael ; James, David E. ; Daly, Roger J. / Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein. In: EMBO Journal. 2008 ; Vol. 27, No. 17. pp. 2305-2316.
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    title = "Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein",
    abstract = "Grb2-associated binder (Gab)2 functions downstream of a variety of receptor and cytoplasmic tyrosine kinases as a docking platform for specific signal transducers and performs important functions in both normal physiology and oncogenesis. Gab2 signalling is promoted by its association with specific receptors through the adaptor Grb2. However, the molecular mechanisms that attenuate Gab2 signals have remained unclear. We now demonstrate that growth factor-induced phosphorylation of Gab2 on two residues, S210 and T391, leads to recruitment of 14-3-3 proteins. Together, these events mediate negative-feedback regulation, as Gab2(S210A/T391A) exhibits sustained receptor association and signalling and promotes cell proliferation and transformation. Importantly, introduction of constitutive 14-3-3-binding sites into Gab2 renders it refractory to receptor activation, demonstrating that site-selective binding of 14-3-3 proteins is sufficient to terminate Gab2 signalling. Furthermore, this is associated with reduced binding of Grb2. This leads to a model where signal attenuation occurs because 14-3-3 promotes dissociation of Gab2 from Grb2, and thereby uncouples Gab2 from the receptor complex. This represents a novel regulatory mechanism with implications for diverse tyrosine kinase signalling systems.",
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    Brummer, T, Larance, M, Herrera Abreu, MT, Lyons, RJ, Timpson, P, Emmerich, CH, Fleuren, EDG, Lehrbach, GM, Schramek, D, Guilhaus, M, James, DE & Daly, RJ 2008, 'Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein', EMBO Journal, vol. 27, no. 17, pp. 2305-2316. https://doi.org/10.1038/emboj.2008.159

    Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein. / Brummer, Tilman; Larance, Mark; Herrera Abreu, Maria Teresa; Lyons, Ruth J.; Timpson, Paul; Emmerich, Christoph H.; Fleuren, Emmy D. G.; Lehrbach, Gillian M.; Schramek, Daniel; Guilhaus, Michael; James, David E.; Daly, Roger J. (Lead / Corresponding author).

    In: EMBO Journal, Vol. 27, No. 17, 03.09.2008, p. 2305-2316.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein

    AU - Brummer, Tilman

    AU - Larance, Mark

    AU - Herrera Abreu, Maria Teresa

    AU - Lyons, Ruth J.

    AU - Timpson, Paul

    AU - Emmerich, Christoph H.

    AU - Fleuren, Emmy D. G.

    AU - Lehrbach, Gillian M.

    AU - Schramek, Daniel

    AU - Guilhaus, Michael

    AU - James, David E.

    AU - Daly, Roger J.

    PY - 2008/9/3

    Y1 - 2008/9/3

    N2 - Grb2-associated binder (Gab)2 functions downstream of a variety of receptor and cytoplasmic tyrosine kinases as a docking platform for specific signal transducers and performs important functions in both normal physiology and oncogenesis. Gab2 signalling is promoted by its association with specific receptors through the adaptor Grb2. However, the molecular mechanisms that attenuate Gab2 signals have remained unclear. We now demonstrate that growth factor-induced phosphorylation of Gab2 on two residues, S210 and T391, leads to recruitment of 14-3-3 proteins. Together, these events mediate negative-feedback regulation, as Gab2(S210A/T391A) exhibits sustained receptor association and signalling and promotes cell proliferation and transformation. Importantly, introduction of constitutive 14-3-3-binding sites into Gab2 renders it refractory to receptor activation, demonstrating that site-selective binding of 14-3-3 proteins is sufficient to terminate Gab2 signalling. Furthermore, this is associated with reduced binding of Grb2. This leads to a model where signal attenuation occurs because 14-3-3 promotes dissociation of Gab2 from Grb2, and thereby uncouples Gab2 from the receptor complex. This represents a novel regulatory mechanism with implications for diverse tyrosine kinase signalling systems.

    AB - Grb2-associated binder (Gab)2 functions downstream of a variety of receptor and cytoplasmic tyrosine kinases as a docking platform for specific signal transducers and performs important functions in both normal physiology and oncogenesis. Gab2 signalling is promoted by its association with specific receptors through the adaptor Grb2. However, the molecular mechanisms that attenuate Gab2 signals have remained unclear. We now demonstrate that growth factor-induced phosphorylation of Gab2 on two residues, S210 and T391, leads to recruitment of 14-3-3 proteins. Together, these events mediate negative-feedback regulation, as Gab2(S210A/T391A) exhibits sustained receptor association and signalling and promotes cell proliferation and transformation. Importantly, introduction of constitutive 14-3-3-binding sites into Gab2 renders it refractory to receptor activation, demonstrating that site-selective binding of 14-3-3 proteins is sufficient to terminate Gab2 signalling. Furthermore, this is associated with reduced binding of Grb2. This leads to a model where signal attenuation occurs because 14-3-3 promotes dissociation of Gab2 from Grb2, and thereby uncouples Gab2 from the receptor complex. This represents a novel regulatory mechanism with implications for diverse tyrosine kinase signalling systems.

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    KW - Recombinant Proteins

    KW - Humans

    KW - Amino Acid Sequence

    KW - Feedback, Physiological

    KW - Models, Biological

    KW - Protein Binding

    KW - Binding Sites

    KW - Mutagenesis, Site-Directed

    KW - Multiprotein Complexes

    KW - Phosphorylation

    KW - Adaptor Proteins, Signal Transducing

    KW - Molecular Sequence Data

    KW - Sequence Homology, Amino Acid

    KW - Signal Transduction

    KW - Cell Line

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    M3 - Article

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    JF - EMBO Journal

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    Brummer T, Larance M, Herrera Abreu MT, Lyons RJ, Timpson P, Emmerich CH et al. Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein. EMBO Journal. 2008 Sep 3;27(17):2305-2316. https://doi.org/10.1038/emboj.2008.159