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Regulation of the Raf–MEK–ERK pathway by protein phosphatase 5

Nature Cell Biology volume 8, pages 1011–1016 (2006)Cite this article

Abstract

The Raf–MEK–ERK pathway couples growth factor, mitogenic and extracellular matrix signals to cell fate decisions such as growth, proliferation, migration, differentiation and survival1,2. Raf-1 is a direct effector of the Ras GTPase and is the initiating kinase in this signalling cascade. Although Raf-1 activation is well studied, little is known about how Raf-1 is inactivated. Here, we used a proteomic approach to identify molecules that may inactivate Raf-1 signalling. Protein phosphatase 5 (PP5) was identified as an inactivator that associates with Raf-1 on growth factor stimulation and selectively dephosphorylates an essential activating site, Ser 338. The PP5-mediated dephosphorylation of Ser 338 inhibited Raf-1 activity and downstream signalling to MEK, an effect that was prevented by phosphomimetic substitution of Ser 338, or by ablation of PP5 catalytic function. Furthermore, depletion of endogenous PP5 increased cellular phospho-Ser 338 levels. Our results suggest that PP5 is a physiological regulator of Raf-1 signalling pathways.

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Figure 1: Interaction between Raf-1 and PP5.
Figure 2: PP5 inhibits Raf-1 activation.
Figure 3: PP5 inhibits Raf-1 signalling by specifically dephosphorylating Ser 338.
Figure 4: PP5 inhibits Raf-1-induced neuritogenesis in PC12 cells in a Ser 338-dependent manner.

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Acknowledgements

We thank D. Morrison, P. Cohen, M. Chinkers and X.-F. Wang for reagents and R. Honkanen for advice on PP5 siRNA. We also thank V. Cleghon for comments on the manuscript. This work was supported by Cancer Research UK, the European Union (FP6 Interaction Proteome) and the Wellcome Trust.

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Authors and Affiliations

  1. The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, G61 1BD, Glasgow, UK

    Alex von Kriegsheim, G. Joan Grindlay, Walter Kolch & Amardeep S. Dhillon

  2. Sir Henry Wellcome Functional Genomics Facility, University of Glasgow, Glasgow, G12 8QQ, UK

    Andrew Pitt & Walter Kolch

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  1. Alex von Kriegsheim
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  2. Andrew Pitt
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  5. Amardeep S. Dhillon
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Correspondence to Walter Kolch or Amardeep S. Dhillon.

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Supplementary Figures S1, S2, S3, S4 and Methods (PDF 416 kb)

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von Kriegsheim, A., Pitt, A., Grindlay, G. et al. Regulation of the Raf–MEK–ERK pathway by protein phosphatase 5. Nat Cell Biol 8, 1011–1016 (2006). https://doi.org/10.1038/ncb1465

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