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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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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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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DOI: https://doi.org/10.1038/ncb783
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