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Originally published In Press as doi:10.1074/jbc.M004861200 on September 19, 2000

J. Biol. Chem., Vol. 275, Issue 49, 38363-38370, December 8, 2000
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Smad7 Is Induced by CD40 and Protects WEHI 231 B-lymphocytes from Transforming Growth Factor-beta -induced Growth Inhibition and Apoptosis*

Supriya Patil, Gary M. Wildey, Thomas L. BrownDagger , Lisa Choy§, Rik Derynck§, and Philip H. Howe

From the Department of Cell Biology, The Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195 and § Departments of Growth Development and Anatomy, Programs in Cell Biology and Developmental Biology, University of California at San Francisco, San Francisco, California 94143-0640

Transforming growth factor-beta (TGF-beta ) is a potent inducer of apoptosis in B-lymphocytes and is essential for immune regulation and maintenance of self-tolerance. Here we show that concomitant signaling through CD40 sustains proliferation and rescues the premature B cell line WEHI 231 from both TGF-beta -induced and anti-IgM-induced apoptosis. The anti-apoptotic effect of CD40 is associated with the transcriptional activation of the inhibitory Smad7 protein. The transactivation of Smad7 by CD40 is NFkappa B-dependent in that pharmacological inhibitors of this pathway, N-tosyl-L-phenylalanine chloromethyl ketone and pyrrolidine dithiocarbamate, abrogate CD40-induced Smad7 expression. Ectopic overexpression of Smad7 inhibited Smad2 activation, TGF-beta -mediated growth inhibition, and apoptosis in WEHI 231 cells. Consistent with this result, dominant negative interference with Smad2 and Smad3 function also inhibited TGF-beta -induced apoptosis. The inhibitory effects of Smad7 overexpression were specific to TGF-beta -induced apoptosis and were without effect on anti-IgM-induced cell death. These results suggest a mechanism of suppression of TGF-beta -induced apoptosis by CD40, mediated through activation of NF-kappa B and, consequently, induction of Smad7 expression.

* This work was supported by National Institutes of Health Grants CA80095 (NCI) (to P. H. H.) and P01-HL60231 (project III) (to R. D.) and a postdoctoral fellowship from the American Heart Association (to L. C.).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 Present address: Dept. of Physiology and Biophysics, School of Medicine, MM-12, Wright State University, 3640 Colonel Glenn Highway, Dayton, OH 45435-0001.

To whom correspondence should be addressed: Dept. of Cell Biology, NC1, The Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44195. Tel.: 216- 445-9750; Fax: 216-445-7855; E-mail: howep@ccf.org.

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