Skip to main content

Thank you for visiting nature.com. 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:

EPS8 and E3B1 transduce signals from Ras to Rac

Nature volume 401, pages 290–293 (1999)Cite this article

Abstract

The small guanine nucleotide (GTP)-binding protein Rac regulates mitogen-induced cytoskeletal changes and c-Jun amino-terminal kinase (JNK), and its activity is required for Ras-mediated cell transformation1. Epistatic analysis placed Rac as a key downstream target in Ras signalling2; however, the biochemical mechanism regulating the cross-talk among these small GTP-binding proteins remains to be elucidated. Eps8 (relative molecular mass 97,000) is a substrate of receptors with tyrosine kinase activity3 which binds, through its SH3 domain, to a protein designated E3b1/Abi-1 (refs 4, 5). Here we show that Eps8 and E3b1/Abi-1 participate in the transduction of signals from Ras to Rac, by regulating Rac-specific guanine nucleotide exchange factor (GEF) activities. We also show that Eps8, E3b1 and Sos-1 form a tri-complex in vivo that exhibits Rac-specific GEF activity in vitro. We propose a model in which Eps8 mediates the transfer of signals between Ras and Rac, by forming a complex with E3b1 and Sos-1.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Disruption of eps8 locus abrogates PDGF-induced membrane ruffling.
Figure 2: Epistatic localization of Eps8 in the Ras pathway.
Figure 3: Eps8 activates Rac and a Rac-specific GEF.
Figure 4: Eps8, E3b1 and Sos-1 form a tri-complex with Rac–GEF activity.
Figure 5: Inhibition of ruffling by anti-E3b1 antibodies.

Similar content being viewed by others

References

  1. Van Aelst,L. & D'Souza-Schorey,C. Rho GTPases and signaling networks. Genes Dev. 11, 2295–2322 (1997).

    Article  CAS  Google Scholar 

  2. Hall,A. Rho GTPases and the actin cytoskeleton. Science 279, 509–514 (1998).

    Article  ADS  CAS  Google Scholar 

  3. 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 

  4. 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 

  5. Shi,Y., Alin,K. & Goff,S. P. Abl-interactor-1, a novel SH3 protein binding to the carboxy-terminal portion of the Abl protein, suppresses v-abl transforming activity. Genes Dev. 9, 2583–2597 (1995).

    Article  CAS  Google Scholar 

  6. Rodriguez-Viciana,P., Marte,B. M., Warne,P. H. & Downward,J. Phosphatidylinositol 3′ kinase: one of the effectors of Ras. Phil. Trans. R. Soc. Lond. B Biol. Sci. 351, 225–231 (1996).

    Article  ADS  CAS  Google Scholar 

  7. Han,J. et al. Role of substrates and products of PI 3-kinase in regulating activation of Rac-related guanosine triphosphatases by Vav. Science 279, 558–560 (1998).

    Article  ADS  CAS  Google Scholar 

  8. Nimnual,A. S., Yatsula,B. A. & Bar-Sagi,D. Coupling of Ras and Rac guanosine triphosphatases through the Ras exchanger Sos. Science 279, 560–563 (1998).

    Article  ADS  CAS  Google Scholar 

  9. Ridley, A J., Paterson,H. F., Johnston,C. L., Diekmann,D. & Hall,A. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell 70, 401–410 (1992).

    Article  Google Scholar 

  10. Manser,E., Leung,T., Salihuddin,H., Zhao,Z. S. & Lim,L. A brain serine/threonine protein kinase activated by Cdc42 and Rac1. Nature 367, 40–46 (1994).

    Article  ADS  CAS  Google Scholar 

  11. 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 

  12. Sastry,L. et al. Quantitative analysis of Grb2–Sos1 interaction: the N-terminal SH3 domain of Grb2 mediates affinity. Oncogene 11, 1107–1112 (1995).

    CAS  PubMed  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. Joneson,T., McDonough,M., Bar-Sagi,D. & Van Aelst,L. RAC regulation of actin polymerization and proliferation by a pathway distinct from Jun kinase. Science 274, 1374–1376 (1996). (Published erratum appears in Science 276, 185 (1997).)

    Article  ADS  CAS  Google Scholar 

  15. Lamarche,N. et al. Rac and Cdc42 induce actin polymerization and G1 cell cycle progression independently of p65PAK and the JNK/SAPK MAP kinase cascade. Cell 87, 519–529 (1996).

    Article  CAS  Google Scholar 

  16. Westwick,J. K. et al. Rac regulation of transformation, gene expression, and actin organization by multiple, PAK-independent pathways. Mol. Cell. Biol. 17, 1324–1335 (1997).

    Article  CAS  Google Scholar 

  17. Reif,K., Nobes,C. D., Thomas,G., Hall,A. & Cantrell,D. A. Phosphatidylinositol 3-kinase signals activate a selective subset of Rac/Rho-dependent effector pathways. Curr. Biol. 6, 1445–1455 (1996).

    Article  CAS  Google Scholar 

  18. Lanfrancone,L. et al. Overexpression of Shc proteins potentiates the proliferative response to the granulocyte-macrophage colony-stimulating factor and recruitment of Grb2/SoS and Grb2/p140 complexes to the beta receptor subunit. Oncogene 10, 907–917 (1995). (Published erratum appears in Oncogene 11, 607 (1995).)

    CAS  PubMed  Google Scholar 

  19. Coso,O. A. et al. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signaling pathway. Cell 81, 1137–1146 (1995).

    Article  CAS  Google Scholar 

  20. Leveen,P. et al. Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities. Genes Dev. 8, 1875–1887 (1994).

    Article  CAS  Google Scholar 

  21. Zheng,Y., Hart,M. J. & Cerione,R. A. Guanine nucleotide exchange catalyzed by dbl oncogene product. Methods Enzymol. 256, 77–84 (1995).

    Article  CAS  Google Scholar 

  22. Manser,E. et al. PAK kinases are directly coupled to the PIX family of nucleotide exchange factors. Mol. Cell 1, 183–192 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank J. Downward, E. Van Obberghen-Shilling, S. Meloche, L. B. Areces, A. Mongiovi and L. Lanfrancone for the gift of various plasmids and mutants; E. Frittoli and S. Beckman for technical assistance; and E. Migliaccio and all the members of the Di Fiore and Betsholtz laboratories for discussions. The microinjector Axiovert 100 (Zeiss) was donated by the Lattanzio family.

Author information

Author notes

  1. Giorgio Scita and Johan Nordstrom: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Experimental Oncology, European Institute of Oncology, Via Ripamonti, 435, 20141, Milan, Italy

    Giorgio Scita, Roberta Carbone, Pierluigi Tenca, Giuseppina Giardina & Pier Paolo Di Fiore

  2. Department of Medical Biochemistry, University of Goteborg, Medicinaregatan 13, Box 440, Goteborg, SE 405 30, Sweden

    Johan Nordstrom, Mattias Bjarnegård & Christer Betsholtz

  3. Istituto di Microbiologia, University of Bari, Bari, 70124, Italy

    Pier Paolo Di Fiore

  4. Molecular Signaling Unit, Laboratory of Cellular Development and Oncology, National Institute of Health, 20892, Bethesda, Maryland, USA

    Silvio Gutkind

Authors
  1. Giorgio Scita
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Johan Nordstrom
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Roberta Carbone
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pierluigi Tenca
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Giuseppina Giardina
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Silvio Gutkind
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mattias Bjarnegård
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Christer Betsholtz
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Pier Paolo Di Fiore
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pier Paolo Di Fiore.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Scita, G., Nordstrom, J., Carbone, R. et al. EPS8 and E3B1 transduce signals from Ras to Rac. Nature 401, 290–293 (1999). https://doi.org/10.1038/45822

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/45822

This article is cited by

Comments

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.

Search

Advanced search

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