Letter | Published:

Argos inhibits epidermal growth factor receptor signalling by ligand sequestration

Nature volume 430, pages 10401044 (26 August 2004) | Download Citation

Subjects

Abstract

The epidermal growth factor receptor (EGFR) has critical functions in development and in many human cancers1,2,3. During development, the spatial extent of EGFR signalling is regulated by feedback loops comprising both well-understood activators and less well-characterized inhibitors3,4. In Drosophila melanogaster the secreted protein Argos functions as the only known extracellular inhibitor of EGFR5, with clearly identified roles in multiple stages of development3. Argos is only expressed when the Drosophila EGFR (DER) is activated at high levels6, and downregulates further DER signalling. Although there is ample genetic evidence that Argos inhibits DER activation, the biochemical mechanism has not been established. Here we show that Argos inhibits DER signalling without interacting directly with the receptor, but instead by sequestering the DER-activating ligand Spitz. Argos binds tightly to the EGF motif of Spitz and forms a 1:1 (Spitz:Argos) complex that does not bind DER in vitro or at the cell surface. Our results provide an insight into the mechanism of Argos function, and suggest new strategies for EGFR inhibitor design.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Epidermal growth factor receptor: mechanisms of activation and signalling. Exp. Cell Res. 284, 31–53 (2003)

  2. 2.

    & Untangling the ErbB signalling network. Nature Rev. Mol. Cell Biol. 2, 127–137 (2001)

  3. 3.

    Signaling by the Drosophila epidermal growth factor receptor pathway during development. Exp. Cell Res. 284, 140–149 (2003)

  4. 4.

    , & Computational modeling of the EGF-receptor system: a paradigm for systems biology. Trends Cell Biol. 12, 43–50 (2003)

  5. 5.

    , , , & Inhibition of Drosophila EGF receptor activation by the secreted protein Argos. Nature 376, 699–702 (1995)

  6. 6.

    , , & Argos transcription is induced by the Drosophila EGF receptor pathway to form an inhibitory feedback loop. Development 122, 223–230 (1996)

  7. 7.

    , , & The argos gene encodes a diffusible factor that regulates cell fate decisions in the Drosophila eye. Cell 69, 963–975 (1992)

  8. 8.

    & An autoregulatory cascade of EGF receptor signaling patterns the Drosophila egg. Cell 95, 355–364 (1998)

  9. 9.

    , , & The interaction between the Drosophila secreted protein argos and the epidermal growth factor receptor inhibits dimerization of the receptor and binding of secreted spitz to the receptor. Mol. Cell. Biol. 20, 2098–2107 (2000)

  10. 10.

    & Evidence that Argos is an antagonistic ligand of the EGF receptor. Oncogene 19, 3560–3562 (2000)

  11. 11.

    et al. EGF activates its receptor by removing interactions that autoinhibit ectodomain dimerization. Mol. Cell 11, 507–517 (2003)

  12. 12.

    , & In vivo analysis of Argos structure-function. Sequence requirements for inhibition of the Drosophila epidermal growth factor receptor. J. Biol. Chem. 273, 4275–4281 (1998)

  13. 13.

    Beyond carrier proteins: Specifying the cellular responses to IGF signals: roles of IGF-binding proteins. J. Endocrinol. 175, 41–54 (2002)

  14. 14.

    et al. Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. Nature 420, 636–642 (2002)

  15. 15.

    , , , & The tethered configuration of the EGF receptor extracellular domain exerts only a limited control of receptor function. Proc. Natl Acad. Sci. USA 101, 923–928 (2004)

  16. 16.

    , , & Direct visualization of binding, aggregation, and internalization of insulin and epidermal growth factor on living fibroblastic cells. Proc. Natl Acad. Sci. USA 75, 2659–2663 (1978)

  17. 17.

    , , & Quantitative imaging of lateral ErbB1 receptor signal propagation in the plasma membrane. Science 290, 1567–1570 (2000)

  18. 18.

    , , & Heparan sulfate proteoglycans retain Noggin at the cell surface: a potential mechanism for shaping bone morphogenetic protein gradients. J. Biol. Chem. 277, 2089–2096 (2002)

  19. 19.

    , , & Interactions of heparin/heparan sulfate with proteins: appraisal of structural factors and experimental approaches. Glycobiology 14, 17R–30R (2004)

  20. 20.

    & Heparan sulfate core proteins in cell–cell signaling. Annu. Rev. Genet. 37, 461–484 (2003)

  21. 21.

    , , & Interleukin-1 receptor antagonist: role in biology. Annu. Rev. Immunol. 16, 27–55 (1998)

  22. 22.

    , , , & Decoy receptors: a strategy to regulate inflammatory cytokines and chemokines. Trends Immunol. 22, 328–336 (2001)

  23. 23.

    & Modeling pattern formation: counting to two in the Drosophila egg. Curr. Biol. 12, R493–R495 (2002)

  24. 24.

    , & Modeling and computational analysis of EGF receptor-mediated cell communication in Drosophila oogenesis. Development 129, 2577–2589 (2002)

  25. 25.

    , , & Epidermal growth factor-related peptides and their receptors in human malignancies. Crit. Rev. Oncol. Hematol. 19, 183–232 (1995)

  26. 26.

    , , , & Extracellular domains drive homo- but not hetero-dimerization of erbB receptors. EMBO J. 19, 4632–4643 (2000)

  27. 27.

    , , & Secreted Spitz triggers the DER signaling pathway and is a limiting component in embryonic ventral ectoderm determination. Genes Dev. 9, 1518–1529 (1995)

Download references

Acknowledgements

We thank J. Duffy, K. Ferguson, P. Carroll, G. Van Duyne and members of the Lemmon and Shvartsman laboratories for valuable discussions; T. Schüpbach and N. Perrimon for providing cDNAs. This work was supported by grants from the NIH (to M.A.L.) and NSF (S.Y.S.), by NIH training grant support (to D.E.K. and V.M.N.), and by an NSF graduate research fellowship (G.T.R.).

Author information

Author notes

    • Daryl E. Klein
    •  & Valerie M. Nappi

    These authors contributed equally to this work

Affiliations

  1. Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, 809C Stellar-Chance Laboratories, 422 Curie Boulevard, Philadelphia, Pennsylvania 19104-6059, USA

    • Daryl E. Klein
    • , Valerie M. Nappi
    •  & Mark A. Lemmon
  2. Department of Chemical Engineering and the Lewis-Sigler Institute for Integrative Genomics, Princeton University, Carl Icahn Laboratory, Washington Road, Princeton, New Jersey 08544, USA

    • Gregory T. Reeves
    •  & Stanislav Y. Shvartsman

Authors

  1. Search for Daryl E. Klein in:

  2. Search for Valerie M. Nappi in:

  3. Search for Gregory T. Reeves in:

  4. Search for Stanislav Y. Shvartsman in:

  5. Search for Mark A. Lemmon in:

Competing interests

The authors declare that they have no competing financial interests.

Corresponding author

Correspondence to Mark A. Lemmon.

Supplementary information

Word documents

  1. 1.

    Supplementary Figure 1

    SPR binding curves showing that the Argos/Spitz interaction requires only the EGF motif of Spitz and the C-terminal 225 amino acids of Argos.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature02840

Further reading

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.