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Originally published In Press as doi:10.1074/jbc.M909012199 on May 22, 2000

J. Biol. Chem., Vol. 275, Issue 32, 24945-24952, August 11, 2000
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Mediation by the Protein-tyrosine Kinase Tec of Signaling between the B Cell Antigen Receptor and Dok-1*

Koji YoshidaDagger §, Yoshihiro YamashitaDagger , Akira MiyazatoDagger , Ken-ichi OhyaDagger ||, Akira Kitanaka**, Uichi Ikeda||, Kazuyuki Shimada||, Takeo Yamanaka§, Keiya Ozawa, and Hiroyuki ManoDagger §§

From the Dagger  Division of Functional Genomics, Departments of  Hematology and || Cardiology, Jichi Medical School, Kawachi-gun, Tochigi 329-0498, Japan, § Omiya Medical Center, Omiya-shi, Saitama 330-8503, Japan, and the ** First Department of Internal Medicine, Kagawa Medical University, Kagawa 761-0793, Japan

A variety of growth factor receptors induce the tyrosine phosphorylation of a nonreceptor protein-tyrosine kinase Tec as well as that of a Tec-binding protein of 62 kDa. Given the similarity in properties between this 62-kDa protein and p62Dok-1, the possibility that these two proteins are identical was investigated. Overexpression of a constitutively active form of Tec in a pro-B cell line induced the hyperphosphorylation of endogenous Dok-1. Tec also associated with Dok-1 in a phosphorylation-dependent manner in 293 cells. Tec mediated marked phosphorylation of Dok-1 both in vivo and in vitro, and this effect required both the Tec homology and Src homology 2 domains of Tec in addition to its kinase activity. Expression of Dok-1 in 293 cells induced inhibition of Ras activity, suggesting that Dok-1 is a negative regulator of Ras. In the immature B cell line Ramos, cross-linking of the B cell antigen receptor (BCR) resulted in tyrosine phosphorylation of Dok-1, and this effect was markedly inhibited by expression of dominant negative mutants of Tec. Furthermore, overexpression of Dok-1 inhibited activation of the c-fos promoter induced by stimulation of the BCR. These results suggest that Tec is an important mediator of signaling from the BCR to Dok-1.

* This work was supported in part by grants-in-aid for Scientific Research on Priority Areas from the Ministry of Education, Science, Sports, and Culture of Japan.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: Division of Functional Genomics, Jichi Medical School, 3311-1 Yakushiji, Kawachi-gun, Tochigi 329-0498, Japan. Tel.: 81-285-58-7449; Fax: 81-285-44-7322; E-mail: hmano@jichi.ac.jp.

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