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Sprouty1 and Sprouty2 provide a control mechanism for the Ras/MAPK signalling pathway

Nature Cell Biology volume 4, pages 850–858 (2002)Cite this article

Abstract

Sprouty (Spry) inhibits signalling by receptor tyrosine kinases; however, the molecular mechanism underlying this function has not been defined. Here we show that after stimulation by growth factors Spry1 and Spry2 translocate to the plasma membrane and become phosphorylated on a conserved tyrosine. Next, they bind to the adaptor protein Grb2 and inhibit the recruitment of the Grb2–Sos complex either to the fibroblast growth factor receptor (FGFR) docking adaptor protein FRS2 or to Shp2. Membrane translocation of Spry is necessary for its phosphorylation, which is essential for its inhibitor activity. A tyrosine-phosphorylated octapeptide derived from mouse Spry2 inhibits Grb2 from binding FRS2, Shp2 or mouse Spry2 in vitro and blocks activation of the extracellular-signal-regulated kinase (ERK) in cells stimulated by growth factor. A non-phosphorylated Spry mutant cannot bind Grb2 and acts as a dominant negative, inducing prolonged activation of ERK in response to FGF and promoting the FGF-induced outgrowth of neurites in PC12 cells. Our findings suggest that Spry functions in a negative feedback mechanism in which its inhibitor activity is controlled rapidly and reversibly by post-translational mechanisms.

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Figure 1: Spry binds to Grb2 and colocalizes with Grb2.
Figure 2: Spry inhibits the recruitment of the Grb2–Sos complex to FRS2 or Shp2.
Figure 3: Tyrosine phosphorylation of Spry and its inhibitor activity.
Figure 4: Tyrosine-phosphorylated Spry binds to Grb2.
Figure 5: Phosphorylation of tyrosine is required for Spry inhibitor activity.
Figure 6: Activity of a tyrosine phosphorylated synthetic peptide.
Figure 7: Spry translocation to the plasma membrane is needed for its phosphorylation.
Figure 8: xSpry1Y53F and mSpry2Y55F are dominant-negative mutants.

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Acknowledgements

We thank M. Kasuga for the mouse Grb2 cDNA; S. Kamakura for the mouse Spry2 cDNA; and M. Kusakabe and S. Nishimoto for technical assistance and discussion. This work was supported by grants from the Ministry of Education, Science and Culture of Japan (to E.N).

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  1. Department of Biophysics, Graduate School of Science, Kyoto University, Sakyo-ku, 606-8502, Kyoto, Japan

    Hiroshi Hanafusa, Satoru Torii & Eisuke Nishida

  2. Department of Cell and Developmental Biology, Graduate School of Biostudies, Kyoto University, Sakyo-ku, 606-8502, Kyoto, Japan

    Hiroshi Hanafusa, Satoru Torii, Takayuki Yasunaga & Eisuke Nishida

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Correspondence to Eisuke Nishida.

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Hanafusa, H., Torii, S., Yasunaga, T. et al. Sprouty1 and Sprouty2 provide a control mechanism for the Ras/MAPK signalling pathway. Nat Cell Biol 4, 850–858 (2002). https://doi.org/10.1038/ncb867

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