The endophilin–CIN85–Cbl complex mediates ligand-dependent downregulation of c-Met


Ligand-dependent downregulation of tyrosine kinase receptors is a critical step for modulating their activity. Upon ligand binding, hepatocyte growth factor (HGF) receptor (Met) is polyubiquitinated1 and degraded2; however, the mechanisms underlying HGF receptor endocytosis are not yet known. Here we demonstrate that a complex involving endophilins, CIN85 and Cbl controls this process. Endophilins3 are regulatory components of clathrin-coated vesicle formation. Through their acyl-transferase activity they are thought to modify the membrane phospholipids and induce negative curvature and invagination of the plasma membrane during the early steps of endocytosis4. Furthermore, by means of their Src-homology 3 domains, endophilins are able to bind CIN85, a recently identified protein that interacts with the Cbl proto-oncogene5. Cbl, in turn, binds and ubiquitinates activated HGF receptor, and by recruiting the endophilin–CIN85 complex, it regulates receptor internalization. Inhibition of complex formation is sufficient to block HGF receptor internalization and to enhance HGF-induced signal transduction and biological responses. These data provide further evidence of a relationship between receptor-mediated signalling and endocytosis, and disclose a novel functional role for Cbl in HGF receptor signalling.

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Figure 1: Endophilins interact with CIN85 and Cbl.
Figure 2: Cbl ubiquitinates Met and recruits the CIN85-endophilin complex.
Figure 3: Expression of dominant interfering mutants of Cbl, CIN85 and endophilin impairs HGF receptor internalization and downregulation.
Figure 4: Impairment of Met downregulation enhances signal transduction and biological responses.


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We thank E. Lantelme, C. Giachino and L. Lanzetti for production of anti-endophilin antibodies and for GST–Endo3, GST–Endo1-2-SH3 constructs; L. Tamagnone for ΔPlexin A3 cDNA; P. De Camilli for dynamin cDNA; Y. Yarden for Cbl and ubiquitin plasmids; I. Dikic for CIN85 reagents and Cbl-ΔSH2 plasmid; and W. Y. Langdon for Cbl-70Z and Cbl-480 constructs. We also thank Y. Yarden, L. Tamagnone and our colleagues for discussions; A. Cignetto for secretarial assistance; and E. Wright for editing the manuscript. The technical assistance of L. Palmas, R. Albano and E. Clemente is acknowledged. This work was partly supported by Associazione E. & E. Rulfo per la Genetica Medica to N.M., by Italian Association for Cancer Research to P.M.C. and by MURST COFIN to S.G.

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Correspondence to Silvia Giordano.

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Petrelli, A., Gilestro, G., Lanzardo, S. et al. The endophilin–CIN85–Cbl complex mediates ligand-dependent downregulation of c-Met. Nature 416, 187–190 (2002).

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