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Originally published In Press as doi:10.1074/jbc.M000039200 on June 7, 2000

J. Biol. Chem., Vol. 275, Issue 40, 30765-30773, October 6, 2000
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Requirement of Ras/MAPK Pathway Activation by Transforming Growth Factor beta  for Transforming Growth Factor beta 1 Production in a Smad-dependent Pathway*

Jianbo Yue and Kathleen M. MulderDagger

From the Department of Pharmacology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033

Our previous results have shown that transforming growth factor beta  (TGFbeta ) rapidly activates Ras, as well as both ERKs and SAPKs. In order to address the biological significance of the activation of these pathways by TGFbeta , here we examined the role of the Ras/MAPK pathways and the Smads in TGFbeta 3 induction of TGFbeta 1 expression in untransformed lung and intestinal epithelial cells. Expression of either a dominant-negative mutant of Ras (RasN17) or a dominant-negative mutant of MKK4 (DN MKK4), or addition of the MEK1 inhibitor PD98059, inhibited the ability of TGFbeta 3 to induce AP-1 complex formation at the TGFbeta 1 promoter, and the subsequent induction of TGFbeta 1 mRNA. The primary components present in this TGFbeta 3-inducible AP-1 complex at the TGFbeta 1 promoter were JunD and Fra-2, although c-Jun and FosB were also involved. Furthermore, deletion of the AP-1 site in the TGFbeta 1 promoter or addition of PD98059 inhibited the ability of TGFbeta 3 to stimulate TGFbeta 1 promoter activity. Collectively, our data demonstrate that TGFbeta 3 induction of TGFbeta 1 is mediated through a signaling cascade consisting of Ras, the MAPKKs MKK4 and MEK1, the MAPKs SAPKs and ERKs, and the specific AP-1 proteins Fra-2 and JunD. Although Smad3 and Smad4 were not detectable in TGFbeta 3-inducible AP-1 complexes at the TGFbeta 1 promoter, stable expression of dominant-negative Smad3 could significantly inhibit the ability of TGFbeta 3 to stimulate TGFbeta 1 promoter activity. Transient expression of dominant-negative Smad4 also inhibited the ability of TGFbeta 3 to transactivate the TGFbeta 1 promoter. Thus, although the Ras/MAPK pathways are essential for TGFbeta 3 induction of TGFbeta 1, Smads may only contribute to this biological response in an indirect manner.

* This work was supported by National Institutes of Health Grants CA51424, CA54816, and CA68444 (to K. M. M.).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.

Dagger To whom correspondence should be addressed: Dept. of Pharmacology H078, Pennsylvania State University College of Medicine, 500 University Dr., Hershey, PA 17033. Tel.: 717-531-6789; Fax: 717-531-5013; E-mail: kmm15@psu.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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