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Phospholipase Cγ activates Ras on the Golgi apparatus by means of RasGRP1

Abstract

Ras proteins regulate cellular growth and differentiation, and are mutated in 30% of cancers. We have shown recently that Ras is activated on and transmits signals from the Golgi apparatus as well as the plasma membrane1,2 but the mechanism of compartmentalized signalling was not determined. Here we show that, in response to Src-dependent activation of phospholipase Cγ1, the Ras guanine nucleotide exchange factor RasGRP1 translocated to the Golgi where it activated Ras. Whereas Ca2+ positively regulated Ras on the Golgi apparatus through RasGRP1, the same second messenger negatively regulated Ras on the plasma membrane by means of the Ras GTPase-activating protein CAPRI3. Ras activation after T-cell receptor stimulation in Jurkat cells, rich in RasGRP1, was limited to the Golgi apparatus through the action of CAPRI, demonstrating unambiguously a physiological role for Ras on Golgi. Activation of Ras on Golgi also induced differentiation of PC12 cells, transformed fibroblasts and mediated radioresistance. Thus, activation of Ras on Golgi has important biological consequences and proceeds through a pathway distinct from the one that activates Ras on the plasma membrane.

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Figure 1: Ha–Ras activation on Golgi and deactivation on plasma membrane requires Src-dependent PLC-γ activity.
Figure 2: RasGRP1 translocates from cytosol to the Golgi.
Figure 3: Ras activation in T cells is restricted to Golgi and is RasGRP1-dependent.
Figure 4: CAPRI deactivates Ras on the plasma membrane.
Figure 5: Activation of Ras on Golgi promotes PC12 neuronal differentiation.

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Acknowledgements

We thank J. Stone for anti-RasGRP1 antibodies, G. Carpenter for PLC-γ-/- cells and E. Skolnik and B. Neel for critically reading the manuscript. This work was supported by grants from the National Institutes of Health (to M.R.P., A.P. and A.D.C), the New York State Breast Cancer Research Program and the Burroughs Wellcome Fund (to M.R.P.), and the Lister Institute (to P.J.C.).

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Correspondence to Mark R. Philips.

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Bivona, T., Pérez de Castro, I., Ahearn, I. et al. Phospholipase Cγ activates Ras on the Golgi apparatus by means of RasGRP1. Nature 424, 694–698 (2003). https://doi.org/10.1038/nature01806

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