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Ras signalling on the endoplasmic reticulum and the Golgi

Nature Cell Biology volume 4, pages 343–350 (2002)Cite this article

Abstract

Current models evoke the plasma membrane (PM) as the exclusive platform from which Ras regulates signalling. We developed a fluorescent probe that reports where and when Ras is activated in living cells. We show that oncogenic H-Ras and N-Ras engage Raf-1 on the Golgi and that endogenous Ras and unpalmitoylated H-Ras are activated in response to mitogens on the Golgi and endoplasmic reticulum (ER), respectively. We also demonstrate that H-Ras that is restricted to the ER can activate the Erk pathway and transform fibroblasts, and that Ras localized on different membrane compartments differentially engages various signalling pathways. Thus, Ras signalling is not limited to the PM, but also proceeds on the endomembrane.

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Figure 1: GFP–Raf-1–RBD reports the activation of Ras on the internal membranes of living cells.
Figure 2: Mitogen-stimulated activation of H-Ras on the PM and Golgi.
Figure 3: Bystander FRET demonstrates mitogen-stimulated activation of endogenous Ras on the PM and Golgi.
Figure 4: Activation of palmitoylation-deficient H-Ras on ER and Golgi membranes in response to growth factors.
Figure 5: Endocytosis is not required for growth factor stimulation of endomembrane-associated Ras.
Figure 6: Src is required for Ras activation on the Golgi and deactivation at the PM.
Figure 7: Activation of palmitoylation-deficient H-Ras stimulates the MAPK pathway.
Figure 8: Activated H-Ras restricted to the ER differentially stimulates signalling pathways and transforms fibroblasts.

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Acknowledgements

We thank P. DeCamilli, C. Hauser, J. Lippincott-Schwartz, D. Littman and C. Machamer for plasmids, A. Kenworthy for insightful discussions and M. Feoktistov for her excellent technical assistance. We also thank the late and ever-inspiring T. Morimoto for his assistance, advice and unwavering encouragement. This work was supported by National Institutes of Health grants AI36224 and GM55279 and by the Burroughs Wellcome Fund to M.R.P., by NIH grants CA67771 and CA76092 to A.D.C. and a General Clinical Research Center NIH grant, NCRR (M01RR00096).

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  1. Vi K. Chiu and Trever Bivona: These authors contributed equally to the work.

Authors and Affiliations

  1. Departments of Medicine, Cell Biology and Pharmacology, New York University School of Medicine, 550 First Avenue, New York, 10016, NY, USA

    Vi K. Chiu, Trever Bivona, Angela Hach, J. Bernard Sajous, Joseph Silletti, Heidi Wiener & Mark R. Philips

  2. Departments of Radiation Oncology and Pharmacology, University of North Carolina School of Medicine, 101 Manning Drive, Chapel Hill, 275997512, NC, USA

    Ronald L. Johnson II & Adrienne D. Cox

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41556_2002_BFncb783_MOESM1_ESM.mov

Movie 1 Rapid Activation of Palmitoylation-deficient H-Ras on Endomembrane in Response to EGF Receptor Stimulation. (MOV 1480 kb)

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Chiu, V., Bivona, T., Hach, A. et al. Ras signalling on the endoplasmic reticulum and the Golgi. Nat Cell Biol 4, 343–350 (2002). https://doi.org/10.1038/ncb783

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