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Mol. Cell. Biol., 08 1997, 4509-4516, Vol 17, No. 8
Copyright © 1997, American Society for Microbiology

Phosphorylation of Raf-1 serine 338-serine 339 is an essential regulatory event for Ras-dependent activation and biological signaling

B Diaz, D Barnard, A Filson, S MacDonald, A King and M Marshall
Department of Medicine, Walther Oncology Center, Indiana University School of Medicine, Indianapolis 46202, USA.

Activation of the Raf serine/threonine protein kinases is tightly regulated by multiple phosphorylation events. Phosphorylation of either tyrosine 340 or 341 in the catalytic domain of Raf-1 has been previously shown to induce the ability of the protein kinase to phosphorylate MEK. By using a combination of mitogenic and enzymatic assays, we found that phosphorylation of the adjacent residue, serine 338, and, to a lesser extent, serine 339 is essential for the biological and enzymatic activities of Raf-1. Replacement of S338 with alanine blocked the ability of prenylated Raf-CX to transform Rat-1 fibroblasts. Similarly, the loss of S338-S339 in Raf-1 prevented protein kinase activation in COS-7 cells by either oncogenic Ras[V12] or v-Src. Consistent with phosphorylation of S338-S339, acidic amino acid substitutions of these residues partially restored transforming activity to Raf-CX, as well as kinase activation of Raf-1 by Ras[V12] or v-Src. Two-dimensional phosphopeptide mapping of wild-type Raf-CX and Raf-CX[A338A339] confirmed the presence of a phosphoserine- containing peptide with the predicted mobility in the wild-type protein which was absent from the mutant. This peptide could be quantitatively precipitated by an antipeptide antibody specific for the 18-residue tryptic peptide containing S338-S339 and was demonstrated to contain only phosphoserine. Phosphorylation of this peptide in Raf-1 was significantly increased by coexpression with Ras[V12]. These data demonstrate that Raf-1 residues 338 to 341 constitute a unique phosphoregulatory site in which the phosphorylation of serine and tyrosine residues contributes to the regulation of Raf by Ras, Src, and Ras-independent membrane localization.


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  • Laird, A. D., Morrison, D. K., Shalloway, D. (1999). Characterization of Raf-1 Activation in Mitosis. J. Biol. Chem. 274: 4430-4439 [Abstract] [Full Text]  
  • Florio, T., Yao, H., Carey, K. D., Dillon, T. J., Stork, P. J. S. (1999). Somatostatin Activation of Mitogen-Activated Protein Kinase via Somatostatin Receptor 1 (SSTR1). Mol. Endocrinol. 13: 24-37 [Abstract] [Full Text]  
  • Papin, C., Denouel-Galy, A., Laugier, D., Calothy, G., Eychene, A. (1998). Modulation of Kinase Activity and Oncogenic Properties by Alternative Splicing Reveals a Novel Regulatory Mechanism for B-Raf. J. Biol. Chem. 273: 24939-24947 [Abstract] [Full Text]  
  • Winkler, D. G., Cutler Jr., R. E., Drugan, J. K., Campbell, S., Morrison, D. K., Cooper, J. A. (1998). Identification of Residues in the Cysteine-rich Domain of Raf-1 That Control Ras Binding and Raf-1 Activity. J. Biol. Chem. 273: 21578-21584 [Abstract] [Full Text]  
  • Cutler, R. E. Jr., Stephens, R. M., Saracino, M. R., Morrison, D. K. (1998). Autoregulation of the Raf-1 serine/threonine kinase. Proc. Natl. Acad. Sci. USA 95: 9214-9219 [Abstract] [Full Text]  
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  • Li, W., Chong, H., Guan, K.-L. (2001). Function of the Rho Family GTPases in Ras-stimulated Raf Activation. J. Biol. Chem. 276: 34728-34737 [Abstract] [Full Text]  
  • Sidovar, M. F., Kozlowski, P., Lee, J. W., Collins, M. A., He, Y., Graves, L. M. (2000). Phosphorylation of Serine 43 Is Not Required for Inhibition of c-Raf Kinase by the cAMP-dependent Protein Kinase. J. Biol. Chem. 275: 28688-28694 [Abstract] [Full Text]  


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