Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

The Eps8 protein coordinates EGF receptor signalling through Rac and trafficking through Rab5


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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: RN-tre is a Rab5-GAP whose catalytic activity is encoded by the TrH domain.
Figure 2: RN-tre is a Rab5-GAP in vivo and its activity is negatively regulated by EGFR.
Figure 3: Binding to Eps8 regulates RN-tre in vivo.
Figure 4: Binding of RN-tre to Eps8 results in the attenuation of Rac-mediated signalling.

Similar content being viewed by others


  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 

Download references


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).

Author information

Authors and Affiliations


Corresponding author

Correspondence to Pier Paolo Di Fiore.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

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).

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI:

This article is cited by


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing