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Originally published In Press as doi:10.1074/jbc.M008870200 on March 13, 2001

J. Biol. Chem., Vol. 276, Issue 21, 18169-18177, May 25, 2001
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Identification of the Mechanisms Regulating the Differential Activation of the MAPK Cascade by Epidermal Growth Factor and Nerve Growth Factor in PC12 Cells*

Shih-chu KaoDagger , Rama K. Jaiswal§, Walter Kolch, and Gary E. LandrethDagger ||

From the Dagger  Department of Neurosciences and the Alzheimer Research Laboratory, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, the § NINDS, National Institutes of Health, Bethesda, Maryland 20892, and  The Beatson Institute for Cancer Research, CRC Beatson Laboratories, Bearsden, Glasgow G61 1BD, United Kingdom

In PC12 cells, epidermal growth factor (EGF) transiently stimulates the mitogen-activated protein (MAP) kinases, ERK1 and ERK2, and provokes cellular proliferation. In contrast, nerve growth factor (NGF) stimulation leads to the sustained activation of the MAPKs and subsequently to neuronal differentiation. It has been shown that both the magnitude and longevity of MAPK activation governs the nature of the cellular response. The activations of MAPKs are dependent upon two distinct small G-proteins, Ras and Rap1, that link the growth factor receptors to the MAPK cascade by activating c-Raf and B-Raf, respectively. We found that Ras was transiently stimulated upon both EGF and NGF treatment of PC12 cells. However, EGF transiently activated Rap1, whereas NGF stimulated prolonged Rap1 activation. The activation of the ERKs was due almost exclusively (>90%) to the action of B-Raf. The transient activation of the MAPKs by EGF was a consequence of the formation of a short lived complex assembling on the EGF receptor itself, composed of Crk, C3G, Rap1, and B-Raf. In contrast, NGF stimulation of the cells resulted in the phosphorylation of FRS2. FRS2 scaffolded the assembly of a stable complex of Crk, C3G, Rap1, and B-Raf resulting in the prolonged activation of the MAPKs. Together, these data provide a signaling link between growth factor receptors and MAPK activation and a mechanistic explanation of the differential MAPK kinetics exhibited by these growth factors.

* This work was supported by Grant IBN10433 from the National Science Foundation (to G. E. L.) and Cancer Research Campaign (to W. K.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| To whom correspondence should be addressed: Alzheimer Research Laboratory, Case Western Reserve University School of Medicine, E504, 10900 Euclid Ave., Cleveland, OH 44106. Tel.: 216-368-6101; Fax: 216-368-3079; E-mail: gel2@po.cwru.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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