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Spred is a Sprouty-related suppressor of Ras signalling


Cellular proliferation, and differentiation of cells in response to extracellular signals, are controlled by the signal transduction pathway of Ras, Raf and MAP (mitogen-activated protein) kinase. The mechanisms that regulate this pathway are not well known. Here we describe two structurally similar tyrosine kinase substrates, Spred-1 and Spred-2. These two proteins contain a cysteine-rich domain related to Sprouty (the SPR domain) at the carboxy terminus. In Drosophila, Sprouty inhibits the signalling by receptors of fibroblast growth factor (FGF) and epidermal growth factor (EGF) by suppressing the MAP kinase pathway2,3,4,5,6,7. Like Sprouty, Spred inhibited growth-factor-mediated activation of MAP kinase. The Ras–MAP kinase pathway is essential in the differentiation of neuronal cells and myocytes. Expression of a dominant negative form of Spred and Spred-antibody microinjection revealed that endogenous Spred regulates differentiation in these types of cells. Spred constitutively associated with Ras but did not prevent activation of Ras or membrane translocation of Raf. Instead, Spred inhibited the activation of MAP kinase by suppressing phosphorylation and activation of Raf. Spred may represent a class of proteins that modulate Ras–Raf interaction and MAP kinase signalling.

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Figure 1: Characterization of Spred molecules.
Figure 2: Effects of Spred on differentiation and Erk2 activity of PC12 cells.
Figure 3: Effects of wild-type (WT) and C-terminal-truncated (ΔC) Spred proteins on differentiation of C2C12 cells and Erk2 activation.
Figure 4: Spred inhibits the activation of MAP kinase by suppressing Raf activation.
Figure 5: Interaction of Spred with Ras and Raf.


  1. Prehoda, K. E., Lee, D. J. & Lim, W. A. Structure of the enabled/VASP homology 1 domain–peptide complex: a key component in the spatial control of actin assembly. Cell 97, 471–480 (1999).

    Article  CAS  Google Scholar 

  2. Hacohen, N., Kramer, S., Sutherland, D., Hiromi, Y. & Krasnow, M. A. sprouty encodes a novel antagonist of FGF signaling that patterns apical branching of the Drosophila airways. Cell 92, 253–263 (1998).

    Article  CAS  Google Scholar 

  3. Casci, T., Vinos, J. & Freeman, M. Sprouty, an intracellular inhibitor of Ras signaling. Cell 96, 655–665 (1999).

    Article  CAS  Google Scholar 

  4. de Maximy, A. A. et al. Cloning and expression pattern of a mouse homologue of Drosophila sprouty in the mouse embryo. Mech. Dev. 81, 213–216 (1999).

    Article  CAS  Google Scholar 

  5. Tefft, J. D. et al. Conserved function of mSpry-2, a murine homolog of Drosophila sprouty, which negatively modulates respiratory organogenesis. Curr. Biol. 9, 219–222 (1999).

    Article  CAS  Google Scholar 

  6. Minowada, G. et al. Vertebrate Sprouty genes are induced by FGF signaling and can cause chondrodysplasia when overexpressed. Development 126, 4465–4475 (1999).

    Article  CAS  Google Scholar 

  7. Reich, A., Sapir, A. & Shilo, B. Sprouty is a general inhibitor of receptor tyrosine kinase signaling. Development 126, 4139–4147 (1999).

    Article  CAS  Google Scholar 

  8. Marshall, C. J. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80, 179–185 (1995).

    Article  CAS  Google Scholar 

  9. Bennett, A. M. & Tonks, N. K. Regulation of distinct stages of skeletal muscle differentiation by mitogen-activated protein kinases. Science 278, 1288–1291 (1997).

    Article  ADS  CAS  Google Scholar 

  10. Wu, Z. et al. p38 and extracellular signal-regulated kinases regulate the myogenic program at multiple steps. Mol. Cell. Biol. 20, 3951–3964 (2000).

    Article  CAS  Google Scholar 

  11. Gille, H. et al. ERK phosphorylation potentiates Elk-1-mediated ternary complex formation and transactivation. EMBO J. 14, 951–962 (1995).

    Article  CAS  Google Scholar 

  12. Yeung, K. et al. Suppression of Raf-1 kinase activity and MAP kinase signalling by RKIP. Nature 401, 173–177 (1999).

    Article  ADS  CAS  Google Scholar 

  13. Mason, C. S. et al. Serine and tyrosine phosphorylations cooperate in Raf, but not B-Raf activation. EMBO J. 18, 2137–2148 (1999).

    Article  CAS  Google Scholar 

  14. King, A. J. et al. The protein kinase Pak3 positively regulates Raf-1 activity through phosphorylation of serine 338. Nature 396, 180–183 (1998).

    Article  ADS  CAS  Google Scholar 

  15. Ridley, A. J. et al. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell 70, 401–410 (1992).

    Article  CAS  Google Scholar 

  16. DeMille, M. M., Kimmel, B. E. & Rubin, G. M. A Drosophila gene regulated by rough and glass shows similarity to ena and VASP. Gene 183, 103–108 (1996).

    Article  CAS  Google Scholar 

  17. Elledge, S. J., Mulligan, J. T., Ramer, S. W., Spottswood, M. & Davis, R. W. Lambda YES: A multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. Proc. Natl Acad. Sci. USA 88, 1731–1735 (1991).

    Article  ADS  CAS  Google Scholar 

  18. Yokouchi, M. et al. Cloning and characterization of APS, an adaptor molecule containing PH and SH2 domains that is tyrosine phosphorylated upon B-cell receptor stimulation. Oncogene 15, 7–15 (1997).

    Article  CAS  Google Scholar 

  19. Yasukawa, H. et al. The JAK-binding protein JAB inhibits Janus tyrosine kinase activity through binding in the activation loop. EMBO J. 18, 1309–1320 (1999).

    Article  CAS  Google Scholar 

  20. Nosaka, T. et al. STAT5 as a molecular regulator of proliferation, differentiation and apoptosis in hematopoietic cells. EMBO J. 18, 4754–4765 (1999).

    Article  CAS  Google Scholar 

  21. Rose, D. W., McCabe, G., Feramisco, J. R. & Adler, M. Expression of c-fos and AP-1 activity in senescent human fibroblasts is not sufficient for DNA synthesis. J. Cell Biol. 119, 1405–1411 (1992).

    Article  CAS  Google Scholar 

  22. Suzuki, A. et al. CIS3/SOCS3/SSI3 plays a negative regulatory role in STAT3 activation and intestinal inflammation. J. Exp. Med. 193, 471–481 (2001).

    Article  CAS  Google Scholar 

  23. Warne, P. H., Viciana, P. R. & Downward, J. Direct interaction of Ras and the amino-terminal region of Raf in vitro. Nature 364, 352–355 (1993).

    Article  ADS  CAS  Google Scholar 

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The first two authors contributed almost equally to this work. We thank H. Ohgusu, M. Sasaki and N. Tanaka for technical assistance; T. Sato and Y. Kaziro for wild-type and mutant Ras; H. Miki for the Drosophila cDNA library; Y. Hiromi for Drosophila Sprouty cDNA; P. Chambon, R. Denton, M. Yokouchi and J.-M. Garnier for library construction; Y. Sako and S. Minoguchi for valuable discussions; and M. Ohara for critical comments on the manuscript and for language assistance. This work was supported in part by grants from the Ministry of Education, Science, Technology, Sports, and Culture of Japan, Japan Research Foundation for Clinical Pharmacology, Fukuoka Cancer Society, Uehara Memorial Foundation and the Mitsubishi Foundation. Murine Spred-1 and Spred-2 are deposited in the nucleotide sequence databases under accession numbers AB063495 and AB063496, respectively.

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Correspondence to Akihiko Yoshimura.

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Wakioka, T., Sasaki, A., Kato, R. et al. Spred is a Sprouty-related suppressor of Ras signalling. Nature 412, 647–651 (2001).

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