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The protein kinase Pak3 positively regulates Raf-1 activity through phosphorylation of serine 338

Nature volume 396, pages 180–183 (1998)Cite this article

A Corrigendum to this article was published on 27 July 2000

Abstract

The pathway involving the signalling protein p21Ras propagates a range of extracellular signals from receptors on the cell membrane to the cytoplasm and nucleus1. The Ras proteins regulate many effectors, including members of the Raf family of protein kinases. Ras-dependent activation of Raf-1 at the plasma membrane involves phosphorylation events, protein–protein interactions and structural changes2,3,4,5,6,7,8. Phosphorylation of serine residues 338 or 339 in the catalytic domain of Raf-1 regulates its activation in response to Ras, Src and epidermal growth factor9,10. Here we show that the p21-activated protein kinase Pak3 phosphorylates Raf-1 on serine 338 in vitro and in vivo. The p21-activated protein kinases are regulated by the Rho-family GTPases Rac and Cdc42 (ref. 11). Our results indicate that signal transduction through Raf-1 depends on both Ras and the activation of the Pak pathway. As guanine-nucleotide-exchange activity on Rac can be stimulated by a Ras-dependent phosphatidylinositol-3-OH kinase12,13, a mechanism could exist through which one Ras effector pathway can be influenced by another.

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Figure 1: Raf-1 S338/339 kinase was purified from rat spleen by sequential chromatography and identified as a specific Raf-1 S338/339 kinase.
Figure 2: Tryptic peptide microsequencing of p37.
Figure 3: Immunoprecipitated Pak3 and recombinant Pak3 phosphorylated the Raf-1 peptide and Raf-CR3.
Figure 4: The effect of Pak3 on Raf-1 activation in vivo in COS-7 cells.

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Acknowledgements

We thank W. S. Lane and the Harvard Microchemistry Facility for tryptic peptide microsequence analysis; R. Wireman for technical assistance; and J. Avruch, R. Cerione and L. Quilliam for discussions. This work was supported by grants from the American Cancer Society and Eli Lilly and Company.

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Authors and Affiliations

  1. Department of Medicine, Division of Hematology/Oncology, Indiana University School of Medicine, Indianapolis, 46202, Indiana, USA

    Alastair J. King, Bruce Diaz, Darlene Barnard & Mark S. Marshall

  2. The Walther Oncology Center, Indianapolis, 46202, Indiana, USA

    Alastair J. King, Huaiyu Sun, Bruce Diaz, Darlene Barnard & Mark S. Marshall

  3. Department of Biochemistry & Molecular Biology, Indiana University School of Medicine, Indianapolis, 46202, Indiana, USA

    Huaiyu Sun, Wenyan Miao, Wenyan Miao & Mark S. Marshall

  4. Department of Pharmacology, Section of Molecular & Cell Biology, Cornell University, Ithaca, 14853, New York, USA

    Shubha Bagrodia

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Correspondence to Mark S. Marshall.

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King, A., Sun, H., Diaz, B. et al. The protein kinase Pak3 positively regulates Raf-1 activity through phosphorylation of serine 338. Nature 396, 180–183 (1998). https://doi.org/10.1038/24184

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