The Eps8 protein coordinates EGF receptor signalling through Rac and trafficking through Rab5 (original) (raw)

Nature volume 408, pages 374–377 (2000)Cite this article

Abstract

How epidermal growth factor receptor (EGFR) signalling is linked to EGFR trafficking is largely unknown. Signalling and trafficking involve small GTPases of the Rho and Rab families, respectively. But it remains unknown whether the signalling relying on these two classes of GTPases is integrated, and, if it is, what molecular machinery is involved. Here we report that the protein Eps8 connects these signalling pathways. Eps8 is a substrate of the EGFR1, which is held in a complex with Sos1 by the adaptor protein E3b1 (ref. 2), thereby mediating activation of Rac2. Through its src homology-3 domain, Eps8 interacts with RN-tre3. We show that RN-tre is a Rab5 GTPase-activating protein, whose activity is regulated by the EGFR. By entering in a complex with Eps8, RN-tre acts on Rab5 and inhibits internalization of the EGFR. Furthermore, RN-tre diverts Eps8 from its Rac-activating function, resulting in the attenuation of Rac signalling. Thus, depending on its state of association with E3b1 or RN-tre, Eps8 participates in both EGFR signalling through Rac, and trafficking through Rab5.

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References

  1. Fazioli, F. et al. Eps8, a substrate for the epidermal growth factor receptor kinase, enhances EGF-dependent mitogenic signals. EMBO J. 12, 3799–3808 (1993).
    Article CAS Google Scholar
  2. Scita, G. et al. EPS8 and E3B1 transduce signals from Ras to Rac. Nature 401, 290–293 ( 1999).
    Article ADS CAS Google Scholar
  3. Matoskova, B., Wong, W. T., Nomura, N., Robbins, K. C. & Di Fiore, P. P. RN-tre specifically binds to the SH3 domain of eps8 with high affinity and confers growth advantage to NIH3T3 upon carboxy-terminal truncation. Oncogene 12, 2679– 2688 (1996).
    CAS PubMed Google Scholar
  4. Neuwald, A. F. A shared domain between a spindle assembly checkpoint protein and Ypt/Rab-specific GTPase-activators. Trends Biochem. Sci. 22, 243–244 (1997).
    Article CAS Google Scholar
  5. Albert, S., Will, E. & Gallwitz, D. Identification of the catalytic domains and their functionally critical arginine residues of two yeast GTPase-activating proteins specific for Ypt/Rab transport GTPases. EMBO J. 18, 5216–5225 (1999).
    Article CAS Google Scholar
  6. Simon, I., Zerial, M. & Goody, R. S. Kinetics of interaction of Rab5 and Rab7 with nucleotides and magnesium ions. J. Biol. Chem. 271, 20470–20478 (1996).
    Article CAS Google Scholar
  7. Rybin, V. et al. GTPase activity of Rab5 acts as a timer for endocytic membrane fusion. Nature 383, 266– 269 (1996).
    Article ADS CAS Google Scholar
  8. Scheffzek, K., Ahmadian, M. R. & Wittinghofer, A. GTPase-activating proteins: helping hands to complement an active site. Trends Biochem. Sci. 23, 257–262 (1998).
    Article CAS Google Scholar
  9. Bucci, C. et al. The small GTPase rab5 functions as a regulatory factor in the early endocytic pathway. Cell 70, 715– 728 (1992).
    Article CAS Google Scholar
  10. Gorvel, J. P., Chavrier, P., Zerial, M. & Gruenberg, J. rab5 controls early endosome fusion in vitro. Cell 64, 915–925 (1991).
    Article CAS Google Scholar
  11. McLauchlan, H. et al. A novel role for Rab5-GDI in ligand sequestration into clathrin-coated pits. Curr. Biol. 8, 34– 45 (1998).
    Article CAS Google Scholar
  12. Papini, E. et al. The small GTP binding protein rab7 is essential for cellular vacuolation induced by Helicobacter pylori cytotoxin. EMBO J. 16, 15–24 (1997).
    Article CAS Google Scholar
  13. Kishan, K. V., Scita, G., Wong, W. T., Di Fiore, P. P. & Newcomer, M. E. The SH3 domain of Eps8 exists as a novel intertwined dimer. Nature Struct. Biol. 4, 739– 743 (1997).
    Article CAS Google Scholar
  14. Mongiovi, A. M. et al. A novel peptide–SH3 interaction. EMBO J. 18, 5300–5309 ( 1999).
    Article CAS Google Scholar
  15. Russell, M., Lange-Carter, C. A. & Johnson, G. L. Direct interaction between Ras and the kinase domain of mitogen-activated protein kinase kinase kinase (MEKK1). J. Biol. Chem. 270, 11757–11760 ( 1995).
    Article CAS Google Scholar
  16. Provenzano, C. et al. Eps8, a tyrosine kinase substrate, is recruited to the cell cortex and dynamic F-actin upon cytoskeleton remodeling. Exp. Cell Res. 242, 186–200 ( 1998).
    Article CAS Google Scholar
  17. Castagnino, P. et al. Direct binding of eps8 to the juxtamembrane domain of EGFR is phosphotyrosine- and SH2-independent. Oncogene 10 , 723–729 (1995).
    CAS PubMed Google Scholar
  18. Anderson, R. G. The caveolae membrane system. Annu. Rev. Biochem. 67 , 199–225 (1998).
    Article CAS Google Scholar
  19. Coso, O. A. et al. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signalling pathway. Cell 81 , 1137–1146 (1995).
    Article CAS Google Scholar
  20. Biesova, Z., Piccoli, C. & Wong, W. T. Isolation and characterization of e3B1, an eps8 binding protein that regulates cell growth. Oncogene 14, 233–241 (1997).
    Article CAS Google Scholar
  21. Ridley, A. J. et al. rho family GTPase activating proteins p190, bcr and rhoGAP show distinct specificities in vitro and in vivo. EMBO J. 12, 5151–5160 (1993).
    Article CAS Google Scholar
  22. Horiuchi, H. et al. A novel Rab5 GDP/GTP exchange factor complexed to Rabaptin-5 links nucleotide exchange to effector recruitment and function. Cell 90, 1149–1159 ( 1997).
    Article CAS Google Scholar
  23. Carbone, R. et al. eps15 and eps15R are essential components of the endocytic pathway. Cancer Res. 57, 5498– 5504 (1997).
    CAS PubMed Google Scholar
  24. Sorkin, A., Di Fiore, P. P. & Carpenter, G. The carboxyl terminus of epidermal growth factor receptor/erbB-2 chimerae is internalisation impaired. Oncogene 8, 3021–3028 (1993).
    CAS PubMed Google Scholar

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Acknowledgements

We thank B. Matoskova for reagents and for helpful discussions, and N. Gholami-Shangolabad for technical assistance. This work was supported by grants from Associazione Italiana Ricerca sul Cancro, from the Armenise-Harvard Foundation and from the CNR (Progetto Biotecnologie).

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Author notes

  1. Savvas Christoforidis
    Present address: Laboratory of Biological Chemistry, Medical School University of Ioannina, 45110, Ioannina, Greece

Authors and Affiliations

  1. Department of Experimental Oncology European Institute of Oncology, Milan, 20141, Italy
    Letizia Lanzetti, Maria Grazia Malabarba, Giorgio Scita & Pier Paolo Di Fiore
  2. Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, c/o European Molecular Biology Laboratory (EMBL), Heidelberg, 69117, Germany
    Vladimir Rybin, Savvas Christoforidis & Marino Zerial
  3. IFOM, The FIRC Institute for Molecular Oncology, Milan, 20134, Italy
    Pier Paolo Di Fiore

Authors

  1. Letizia Lanzetti
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  2. Vladimir Rybin
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  3. Maria Grazia Malabarba
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  4. Savvas Christoforidis
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  5. Giorgio Scita
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  6. Marino Zerial
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  7. Pier Paolo Di Fiore
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Correspondence toPier Paolo Di Fiore.

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Lanzetti, L., Rybin, V., Malabarba, M. et al. The Eps8 protein coordinates EGF receptor signalling through Rac and trafficking through Rab5.Nature 408, 374–377 (2000). https://doi.org/10.1038/35042605

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