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Mol. Cell. Biol. 22 (13): 4750-4759

Copyright © 2002 by the American Society for Microbiology. All rights reserved.

Internalization-Dependent and -Independent Requirements for Transforming Growth Factor ß Receptor Signaling via the Smad Pathway

Sumedha G. Penheiter, Hugh Mitchell, Nandor Garamszegi, Maryanne Edens, Jules J. E. Doré, Jr., and Edward B. Leof*

Thoracic Diseases Research Unit and Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905

Received for publication 2 October 2001. Revision received 8 November 2001. Accepted for publication 18 March 2002.

Abstract: Members of the transforming growth factor ß (TGF-ß) family of proteins signal through cell surface transmembrane serine/threonine protein kinases known as type I and type II receptors. The TGF-ß signal is extended through phosphorylation of receptor-associated Smad proteins by the type I receptor. Although numerous investigations have established the sequence of events in TGF-ß receptor (TGF-ßR) activation, none have examined the role of the endocytic pathway in initiation and/or maintenance of the signaling response. In this study we investigated whether TGF-ßR internalization modulates type I receptor activation, the formation of a functional receptor/Smad/SARA complex, Smad2/3 phosphorylation or nuclear translocation, and TGF-ß-dependent reporter gene activity. Our data provide evidence that, whereas type I receptor phosphorylation and association of SARA and Smad2 with the TGF-ßR complex take place independently of clathrin lattice formation, Smad2 or Smad3 activation and downstream signaling only occur after endocytic vesicle formation. Thus, TGF-ßR endocytosis is not simply a way to dampen the signaling response but instead is required to propagate signaling via the Smad pathway.

* Corresponding author. Mailing address: Stabile 858, Mayo Clinic, Rochester, MN 55905. Phone: (507) 284-5717. Fax: (507) 284-4521. E-mail: leof.edward{at}mayo.edu.

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