Protein Arginine Methyltransferase 5 Regulates ERK1/2 Signal Transduction Amplitude and Cell Fate Through CRAF

Arginine methylation of RAF proteins limits ERK activation after growth factor stimulation, thereby determining the biological response. Minimized by Methylation Many growth factors signal through the RAS to RAF to extracellular signal–regulated kinase (RAS-ERK) cascade, a signaling pathway that involves the sequential phosphorylation and activation of a series of protein kinases. Despite their common activation of RAS-ERK signaling, however, different growth factors elicit distinct biological responses. For instance, nerve growth factor (NGF) stimulates differentiation of PC12 cells, whereas epidermal growth factor (EGF) stimulates PC12 cell proliferation. Andreu-Pérez et al. found that the amplitude and duration of ERK phosphorylation in response to certain growth factors are kept in check by methylation of activated RAF proteins, a modification that enhanced RAF protein degradation and thereby limited downstream signaling in the RAS-ERK pathway. When RAF methylation was prevented experimentally, the amplitude and duration of the ERK signal elicited by EGF in PC12 cells (normally smaller and briefer than that elicited by NGF) increased, and the biological response switched from proliferation to differentiation. The RAS to extracellular signal–regulated kinase (ERK) signal transduction cascade is crucial to cell proliferation, differentiation, and survival. Although numerous growth factors activate the RAS-ERK pathway, they can have different effects on the amplitude and duration of the ERK signal and, therefore, on the biological consequences. For instance, nerve growth factor, which elicits a larger and more sustained increase in ERK phosphorylation in PC12 cells than does epidermal growth factor (EGF), stimulates PC12 cell differentiation, whereas EGF stimulates PC12 cell proliferation. Here, we show that protein arginine methylation limits the ERK1/2 signal elicited by particular growth factors in different cell types from various species. We found that this restriction in ERK1/2 phosphorylation depended on methylation of RAF proteins by protein arginine methyltransferase 5 (PRMT5). PRMT5-dependent methylation enhanced the degradation of activated CRAF and BRAF, thereby reducing their catalytic activity. Inhibition of PRMT5 activity or expression of RAF mutants that could not be methylated not only affected the amplitude and duration of ERK phosphorylation in response to growth factors but also redirected the response of PC12 cells to EGF from proliferation to differentiation. This additional level of regulation within the RAS pathway may lead to the identification of new targets for therapeutic intervention.

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