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Phospholipase D2-generated phosphatidic acid couples EGFR stimulation to Ras activation by Sos

Nature Cell Biology volume 9, pages 707–712 (2007)Cite this article

Abstract

The activation of Ras by the guanine nucleotide-exchange factor Son of sevenless (Sos) constitutes the rate-limiting step in the transduction process that links receptor tyrosine kinases to Ras-triggered intracellular signalling pathways. A prerequisite for the function of Sos in this context is its ligand-dependent membrane recruitment, and the prevailing model implicates both the Sos carboxy-terminal proline-rich motifs and amino-terminal pleckstrin homology (PH) domain in this process. Here, we describe a previously unrecognized pathway for the PH domain-dependent membrane recruitment of Sos that is initiated by the growth factor-induced generation of phosphatidic acid via the signalling enzyme phospholipase D2 (PLD2). Phosphatidic acid interacts with a defined site in the Sos PH domain with high affinity and specificity. This interaction is essential for epidermal growth factor (EGF)-induced Sos membrane recruitment and Ras activation. Our findings establish a crucial role for PLD2 in the coupling of extracellular signals to Sos-mediated Ras activation, and provide new insights into the spatial coordination of this activation event.

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Figure 1: The PH domain of Sos (Sos-PH) binds to phosphatidic Acid (PA).
Figure 2: The interaction between Sos-PH and PA is essential for serum-induced Sos membrane recruitment and Sos-mediated Ras activation.
Figure 3: PLD2-mediated signalling is essential for Sos membrane recruitment and Sos-mediated Ras activation.
Figure 4: The transforming activity of Ras is potentiated by PLD2.

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Acknowledgements

The authors thank members of the Bar-Sagi and Frohman laboratories, J. Kuriyan and H. Sondermann for helpful discussion, S. McLaughlin for help with the lipid vesicle binding assay and L. Taylor for help with image capturing and analysis. This work was supported by research grants from National Institutes of Health to D.B.-S. (CA55360 and CA28146), G.D. (GM071475) and M.A.F. (DK64166 and GM71520), and a Scientist Development Grant from the American Heart Association to G.D. (0430096N).

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Authors and Affiliations

  1. Graduate Program in Genetics, Stony Brook University, Stony Brook, 11794, NY, USA

    Chen Zhao, Michael A. Frohman & Dafna Bar-Sagi

  2. Department of Molecular Genetics and Microbiology Stony Brook University, Stony Brook, 11794, NY, USA

    Chen Zhao & Dafna Bar-Sagi

  3. Department of Biochemistry, NYU School of Medicine New York, 10016, NY, USA

    Chen Zhao

  4. Department of Pharmacology and Center for Developmental Genetics, Stony Brook University, Stony Brook, 11794, NY, USA

    Guangwei Du & Michael A. Frohman

  5. Department of Physiology and Biophysics, Stony Brook University, Stony Brook, 11794, NY, USA

    Karl Skowronek & Dafna Bar-Sagi

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Contributions

C.Z. and D.B-S. conceived and designed the experiments. C.Z. performed the experiments. G.D. and M.A.F. designed, generated and provided PLD2 reagents. K.S. performed preliminary lipid-binding experiments.

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Correspondence to Dafna Bar-Sagi.

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The authors declare no competing financial interests.

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Zhao, C., Du, G., Skowronek, K. et al. Phospholipase D2-generated phosphatidic acid couples EGFR stimulation to Ras activation by Sos. Nat Cell Biol 9, 707–712 (2007). https://doi.org/10.1038/ncb1594

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