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Protein kinase Cα activates RAF-1 by direct phosphorylation

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Abstract

THE kinase Raf-1 can be activated by treatment of cells with mitogens and by the protein kinase C (PKC) activator 12-0-tetradecanoyl-phorbol-13-acetate (TPA) (reviewed in refs 1,2). Activated Raf-1 triggers a protein kinase cascade by direct phosphorylation of MAP kinase kinase3–5, resulting in phosphorylation of ternary complex factor6 and Jim7,8 by MAP kinase. Here we investigate the molecular mechanism and biological consequences of PKCα-mediated Raf-1 activation in NIH3T3 fibroblasts. PKCα directly phosphorylates and activates Raf-1 both in vitro and in vivo. PKCα induces Raf-1 phosphorylation at several sites, including a serine residue at position 499. Mutation of serine at this position or at residue 259 does not abrogate Raf-1 stimulation by a combination of Ras plus the src tyrosine kinase Lck, but severely impedes Raf-1 activation by PKCα. Consistent with such a direct interaction is the observation that Raf-1 and PKCα cooperate in the transformation of NIH3T3 cells. The Ser499 phosphorylation site is necessary for this synergism.

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Author notes

  1. Walter Kolch

    Present address: Institute for Clinical Molecular Biology and Tumor Genetics, GSF, Haemato-logikum, Marchioninistrasse 25, D 8000, Muenchen, 70, Germany

Authors and Affiliations

  1. Biological Research, Goedecke AG, Mooswaldallee 1–9, D-7800, Freiburg, Germany

    Walter Kolch, Georg Kochs, Richard Hummel & Dieter Marmé

  2. Laboratory of Viral Carcinogenesis, NIH/NCI, FCRDC, Bldg 560, Frederick, Maryland, 21702, USA

    Gisela Heidecker, Haleh Vahidi & Ulf R. Rapp

  3. Institute of Molecular Cell Biology, University of Freiburg, Goedecke AG, Mooswaldallee 1–9–7800, Freiburg, Germany

    Georg Kochs, Richard Hummel, Günter Finkenzeller & Dieter Marmé

  4. Laboratory of Genetics, NIH/NCI, Bldg 37, Bethesda, Maryland, 20892, USA

    Harald Mischak

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  1. Walter Kolch
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  2. Gisela Heidecker
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  3. Georg Kochs
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  4. Richard Hummel
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  5. Haleh Vahidi
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  6. Harald Mischak
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  7. Günter Finkenzeller
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  8. Dieter Marmé
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  9. Ulf R. Rapp
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Kolch, W., Heidecker, G., Kochs, G. et al. Protein kinase Cα activates RAF-1 by direct phosphorylation. Nature 364, 249–252 (1993). https://doi.org/10.1038/364249a0

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  • DOI: https://doi.org/10.1038/364249a0

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