title: Co-recruitment analysis of the CBL and CBLB signalosomes in primary T cells identifies CD5 as a key regulator of TCR-induced ubiquitylation.
abstract: T-cell receptor (TCR) signaling is essential for the function of T cells and negatively regulated by the E3 ubiquitin-protein ligases CBL and CBLB Here, we combined mouse genetics and affinity purification coupled to quantitative mass spectrometry to monitor the dynamics of the CBL and CBLB signaling complexes that assemble in normal T cells over 600 seconds of TCR stimulation. We identify most previously known CBL and CBLB interacting partners, as well as a majority of proteins that have not yet been implicated in those signaling complexes. We exploit correlations in protein association with CBL and CBLB as a function of time of TCR stimulation for predicting the occurrence of direct physical association between them. By combining co-recruitment analysis with biochemical analysis, we demonstrated that the CD5 transmembrane receptor constitutes a key scaffold for CBL- and CBLB-mediated ubiquitylation following TCR engagement. Our results offer an integrated view of the CBL and CBLB signaling complexes induced by TCR stimulation and provide a molecular basis for their negative regulatory function in normal T cells.
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Voisinne
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France.
authors.forename: Antonio
authors.initials: A
authors.lastname: García-Blesa
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Département Biologie Structural Biophysique, Protéomique Génopole Toulouse Midi Pyrénées, CNRS UMR 5089, Toulouse Cedex, France.
authors.forename: Karima
authors.initials: K
authors.lastname: Chaoui
authors.affiliation: Centre d'Immunophénomique, Aix Marseille Université UM2, Inserm US012, CNRS UMS3367, Marseille, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Fiore
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France.
authors.forename: Elise
authors.initials: E
authors.lastname: Bergot
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France Centre d'Immunophénomique, Aix Marseille Université UM2, Inserm US012, CNRS UMS3367, Marseille, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Girard
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France Centre d'Immunophénomique, Aix Marseille Université UM2, Inserm US012, CNRS UMS3367, Marseille, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Malissen
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Département Biologie Structural Biophysique, Protéomique Génopole Toulouse Midi Pyrénées, CNRS UMR 5089, Toulouse Cedex, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Burlet-Schiltz
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Département Biologie Structural Biophysique, Protéomique Génopole Toulouse Midi Pyrénées, CNRS UMR 5089, Toulouse Cedex, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Gonzalez de Peredo
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France Centre d'Immunophénomique, Aix Marseille Université UM2, Inserm US012, CNRS UMS3367, Marseille, France bernardm@ciml.univ-mrs.fr roncagalli@ciml.univ-mrs.fr.
authors.forename: Bernard
authors.identifier: 0000-0003-1340-9342
authors.initials: B
authors.lastname: Malissen
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille, France bernardm@ciml.univ-mrs.fr roncagalli@ciml.univ-mrs.fr.
authors.forename: Romain
authors.identifier: 0000-0001-7554-0552
authors.initials: R
authors.lastname: Roncagalli
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D018956:CD5 Antigens / Q000378:metabolism*; D053263:Gene Regulatory Networks; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D060066:Protein Interaction Maps; D040901:Proteomics / Q000379:methods*; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism; D054875:Ubiquitination
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018956:CD5 Antigens; C118387:Cblb protein, mouse; C526836:Cd5 protein, mouse; D011948:Receptors, Antigen, T-Cell; D050721:Proto-Oncogene Proteins c-cbl; C493849:Cbl protein, mouse
keywords: CBL; CBLB; CD5; ubiquitylation