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Rap1 mediates sustained MAP kinase activation induced by nerve growth factor

Nature volume 392, pages 622–626 (1998)Cite this article

Abstract

Activation of mitogen-activated protein (MAP) kinase (also known as extracellular-signal-regulated kinase, or ERK)1 by growth factors can trigger either cell growth or differentiation. The intracellular signals that couple growth factors to MAP kinase may determine the different effects of growth factors: for example, transient activation of MAP kinase by epidermal growth factor stimulates proliferation of PC12 cells1, whereas they differentiate in response to nerve growth factor, which acts partly by inducing a sustained activation of MAP kinase1. Here we show that activation of MAP kinase by nerve growth factor involves two distinct pathways: the initial activation of MAP kinase requires the small G protein Ras, but its activation is sustained by the small G protein Rap1. Rap1 is activated by CRK adaptor proteins and the guanine-nucleotide-exchange factor C3G, and forms a stable complex with B-Raf, an activator of MAP kinase. Rap1 is required for at least two indices of neuronal differentiation by nerve growth factor: electrical excitability and the induction of neuron-specific genes. We propose that the activation of Rap1 by C3G represents a common mechanism to induce sustained activation of the MAP kinase cascade in cells that express B-Raf.

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Figure 1: Ras- and Rap-dependent components of NGF-induced activation of ERK.
Figure 2: Rap1 activation and association with B-Raf.
Figure 3: The effect of RapN17 on neuronal differentiation.
Figure 4: Enhancement of NGF signalling by Crk-L and C3G.
Figure 5: Requirement of Crk/C3G for maximal activation of ERK.

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References

  1. Marshall, C. J. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80, 179–185 (1995).

    Article  CAS  Google Scholar 

  2. Dichter, M. A., Tischler, A. S. & Greene, L. A. Nerve growth factor-induced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line. Nature 268, 501–504 (1977).

    Article  ADS  CAS  Google Scholar 

  3. Greenberg, M. E., Greene, L. A. & Ziff, E. B. Nerve growth factor and epidermal growth factor induce rapid transient changes in proto-oncogene transcription in PC12 cells. J. Biol. Chem. 260, 14101–14110 (1958).

    Google Scholar 

  4. Cowley, S., Paterson, H., Kemp, P. & Marshall, C. J. Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH3T3 cells. Cell 77, 841–852 (1994).

    Article  CAS  Google Scholar 

  5. Traverse, S.et al. EGF triggers neuronal differentiation of PC12 cells that overexpress the EGF receptor. Curr. Biol. 4, 694–701 (1994).

    Article  CAS  Google Scholar 

  6. Pan, M.-G., Wang, Y.-H., Hirsch, D. D., Labudda, K. & Stork, P. J. S. The Wnt-1 proto-oncogene regulates MAP kinase activation by multiple growth factors in PC12 cells. Oncogene 11, 2005–2012 (1995).

    CAS  PubMed  Google Scholar 

  7. Yao, H.et al. Cyclic adenosine monophosphate can convert epidermal growth factor into a differentiating factor in neuronal cells. J. Biol. Chem. 270, 20748–20753 (1995).

    Article  CAS  Google Scholar 

  8. Porfiri, E. & McCormick, F. Regulation of epidermal growth factor receptor signaling by phosphorylation of the ras exchange factor hSOS1. J. Biol. Chem. 271, 5871–5877 (1996).

    Article  CAS  Google Scholar 

  9. Vossler, M.et al. cAMP activates MAP kinase and Elk-1 through a B-Raf- and Rap1-dependent pathway. Cell 89, 73–82 (1997).

    Article  CAS  Google Scholar 

  10. Thomas, S. M., DeMarco, M., D'Arcangelo, G., Halegoua, S. & Brugge, J. S. Ras is essential for nerve growth factor- and phorbol ester-induced tyrosine phosphorylation of MAP kinases. Cell 78, 1031–1040 (1992).

    Article  Google Scholar 

  11. Wood, K. W., Sarnecki, C., Roberts, T. M. & Blenis, J. Ras mediates nerve growth factor receptor modulation of three signal-transducing protein kinases: MAP kinase, raf-1, and RSK. Cell 68, 1041–1050 (1992).

    Article  CAS  Google Scholar 

  12. Jaiswal, R. K., Weissinger, E., Kolch, W. & Landreth, G. E. Nerve growth factor-mediated activation of the mitogen-activated protein (MAP) kinase cascade involves a signaling complex containing B-Raf and HSP90. J. Biol. Chem. 271, 23626–23629 (1996).

    Article  CAS  Google Scholar 

  13. Ihara, S.et al. Dual control of neurite outgrowth by STAT3 and MAP kinase in PC12 cells stimulated with interleukin-6. EMBO J. 16, 5345–5352 (1997).

    Article  CAS  Google Scholar 

  14. Mandel, G., Cooperman, S. S., Maue, R. A., Goodman, R. H. & Brehn, P. Selective induction of brain type II Na+ channels by nerve growth factor. Proc. Natl Acad. Sci. 85, 924–928 (1988).

    Article  ADS  CAS  Google Scholar 

  15. Fanger, G. R., Erhardt, P., Cooper, G. M. & Maue, R. A. Ras-independent induction of rat brain type II sodium channel expression in nerve growth factor-treated PC12 cells. J. Neurochem. 61, 1977–1980 (1993).

    Article  CAS  Google Scholar 

  16. D'Arcangelo, G. & Halegoua, S. Abranched signaling pathway for nerve growth factor is revealed by src-, ras-, and raf-mediated gene inductions. Mol. Cell. Biol. 13, 3146–3155 (1993).

    Article  CAS  Google Scholar 

  17. deSouza, S., Lochner, J., Machida, C. M., Matrisian, L. M. & Ciment, G. Anovel NGF-responsive element in the stromelysin-1 (transin) gene that is necessary and sufficient for gene expression in PC12 cells. J. Biol. Chem. 270, 9106–9114 (1995).

    Article  CAS  Google Scholar 

  18. Tanaka, S.et al. C3G, a guanine nucleotide-releasing protein expressed ubiquitously, binds to the Src homology 3 domains of CRK and GRB2/ASH proteins. Proc. Natl Acad. Sci. USA 91, 3443–3447 (1994).

    Article  ADS  CAS  Google Scholar 

  19. Knudsen, B. S., Feller, S. M. & Hanafusa, H. Four proline-rich sequences of the guanine-nucleotide exchange factor C3G bind with unique specificity to the first Src homology 3 domain of Crk. J. Biol. Chem. 269, 32781–32787 (1994).

    CAS  PubMed  Google Scholar 

  20. Gotoh, T.et al. Identification of Rap1 as a target for the Crk SH3 domain-binding guanine nucleotide-releasing factor C3G. Mol. Cell. Biol. 15, 6746–6753 (1995).

    Article  CAS  Google Scholar 

  21. Matsuda, M.et al. CRK protein binds to two guanine nucleotide-releasing proteins for the Ras family and modulates nerve growth factor-induced activation of Ras in PC12 cells. Mol. Cell. Biol. 14, 5495–5500 (1994).

    Article  CAS  Google Scholar 

  22. Feller, S. M., Knudsen, B. & Hanafusa, H. Cellular proteins binding to the first Src homology 3 (SH3) domain of the proto-oncogene product c-Crk indicate Crk-specific signaling pathways. Oncogene 10, 1465–1473 (1995).

    CAS  PubMed  Google Scholar 

  23. Reedquist, K. A.et al. Stimulation through the T cell receptor induces Cbl association with Crk proteins and the guanine nucleotide exchange protein C3G. J. Biol. Chem. 271, 8435–8442 (1996).

    Article  CAS  Google Scholar 

  24. Smit, L., van der Horst, G. & Borst, J. Sos, Vav, and C3G participate in B cell receptor-induced signaling pathways and differentially associate with Shc-Grb2, Crk, and Crk-L adaptors. J. Biol. Chem. 271, 8564–8569 (1996).

    Article  CAS  Google Scholar 

  25. Uemura, N.et al. The BCR/ABL oncogene alters interaction of the adapter proteins Crk-L and Crk with cellular proteins. Leukemia 11, 376–385 (1997).

    Article  CAS  Google Scholar 

  26. Tanaka, S., Ouchi, T. & Hanafusa, H. Downstream of Crk adaptor signaling pathway: activation of jun kinase by v-Crk through the guanine nucleotide exchange protein C3G. Proc. Natl Acad. Sci. USA 94, 2356–2361 (1996).

    Article  ADS  Google Scholar 

  27. Hirsch, D. D. & Stork, P. J. S. MAP kinase phosphatases incativate stress-activated protein kinase pathways in vivo. J. Biol. Chem. 272, 4568–4575 (1997).

    Article  CAS  Google Scholar 

  28. Tanaka, S.et al. Both the SH2 and SH3 domains of human Crk protein are required for neuronal differentiation of PC12 cells. Mol. Cell. Biol. 13, 4409–4415 (1993).

    Article  CAS  Google Scholar 

  29. Tanaka, M., Gupta, R. & Mayer, B. J. Differential inhibition of signaling pathways by dominant-negative SH2/SH3 adaptor proteins. Mol. Cell. Biol. 15, 6829–6837 (1995).

    Article  CAS  Google Scholar 

  30. Boussiotis, V. A., Freeman, G. J., Berezovshaya, A., Barber, D. L. & Nadler, L. M. Maintenance of human T cell anergy: blocking of IL-2 gene transcription by activated Rap1. Science 278, 124–128 (1997).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank C. Marshall, M. Matsuda and B. Druker for cDNAs; S. Grewal, B. Druker and G. Mandel for discussion; and C. Fenner for administrative assistance.

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Authors and Affiliations

  1. The Vollum Institute for Advanced Biomedical Research, Portland, 97201, Oregon, USA

    Randall D. York, Hong Yao, Tara Dillon, Cindy L. Ellig, Stephani P. Eckert & Edwin W. McCleskey

  2. Department of Pathology, Oregon Health Sciences University, Portland, 97201, Oregon, USA

    Philip J. S. Stork

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  1. Randall D. York
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Correspondence to Philip J. S. Stork.

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York, R., Yao, H., Dillon, T. et al. Rap1 mediates sustained MAP kinase activation induced by nerve growth factor. Nature 392, 622–626 (1998). https://doi.org/10.1038/33451

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