HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kesti, T. Articles by Saksela, K. Search for Related Content PubMed PubMed Citation Articles by Kesti, T. Articles by Saksela, K.

Reciprocal Regulation of SH3 and SH2 Domain Binding via Tyrosine Phosphorylation of a Common Site in CD3¹

Tapio Kesti^*, Anja Ruppelt, Jing-Huan Wang, Michael Liss, Ralf Wagner^,¶, Kjetil Taskén and Kalle Saksela^2,*,

^* Department of Virology, Haartman Institute, University of Helsinki and Helsinki University Central Hospital, Helsinki, Finland; Biotechnology Centre, University of Oslo, Oslo, Norway; Institute of Medical Technology, University of Tampere and Tampere University Hospital, Tampere, Finland; Geneart, Regensburg, Germany; and ^¶ Institute of Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany

Recruitment of cellular signaling proteins by the CD3 polypeptides of the TCR complex mediates T cell activation. We have screened a human Src homology 3 (SH3) domain phage display library for proteins that can bind to the proline-rich region of CD3. This screening identified Eps8L1 (epidermal growth factor receptor pathway substrate 8-like 1) together with the N-terminal SH3 domain of Nck1 and Nck2 as its preferred SH3 partners. Studies with recombinant proteins confirmed strong binding of CD3 to Eps8L1 and Nck SH3 domains. CD3 bound well also to Eps8 and Eps8L3, and modestly to Eps8L2, but not detectably to other SH3 domains tested. Interestingly, binding of Nck and Eps8L1 SH3 domains was mapped to a PxxDY motif that shared its tyrosine residue (Y166) with the ITAM of CD3. Phosphorylation of this residue abolished binding of Eps/Nck SH3 domains in peptide spot filter assays, as well as in cells cotransfected with a dominantly active Lck kinase. TCR ligation-induced binding and phosphorylation-dependent loss of binding were also demonstrated between Eps8L1 and endogenous CD3 in Jurkat T cells. Thus, phosphorylation of Y166 serves as a molecular switch during T cell activation that determines the capacity of CD3 to interact with either SH3 or SH2 domain-containing proteins.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by grants from the Academy of Finland, Medical Research Council of Tampere University Hospital, Medical Research Council of Helsinki University Hospital, and Sigrid Juselius Foundation (to K.S.), and from the Functional Genomics Program (FUGE), The Research Council of Norway, Norwegian Cancer Society, and Novo Nordic Foundation Committee (to K.T.).

² Address correspondence and reprint requests to Dr. Kalle Saksela, Department of Virology, Haartman Institute, University of Helsinki, Haartmaninkatu 3, FIN-0014 Helsinki, Finland. E-mail address: kalle.saksela{at}helsinki.fi

³ Abbreviations used in this paper: SH, Src homology domain; TBP, TATA-box binding protein; HA, hemagglutinin.

This article has been cited by other articles:

				J.-F. Brodeur, S. Li, O. Damlaj, and V. P. Dave Expression of fully assembled TCR-CD3 complex on double positive thymocytes: synergistic role for the PRS and ER retention motifs in the intra-cytoplasmic tail of CD3{varepsilon} Int. Immunol., October 9, 2009; (2009) dxp098v1. [Abstract] [Full Text] [PDF]

				J.-F. Brodeur, S. Li, M. d. S. Martins, L. Larose, and V. P. Dave Critical and Multiple Roles for the CD3{epsilon} Intracytoplasmic Tail in Double Negative to Double Positive Thymocyte Differentiation J. Immunol., April 15, 2009; 182(8): 4844 - 4853. [Abstract] [Full Text] [PDF]

				P. Tailor, S. Tsai, A. Shameli, P. Serra, J. Wang, S. Robbins, M. Nagata, A. L. Szymczak-Workman, D. A. A. Vignali, and P. Santamaria The Proline-Rich Sequence of CD3{epsilon} as an Amplifier of Low-Avidity TCR Signaling J. Immunol., July 1, 2008; 181(1): 243 - 255. [Abstract] [Full Text] [PDF]

				L. S. Heikkinen, A. Kazlauskas, K. Melen, R. Wagner, T. Ziegler, I. Julkunen, and K. Saksela Avian and 1918 Spanish Influenza A Virus NS1 Proteins Bind to Crk/CrkL Src Homology 3 Domains to Activate Host Cell Signaling J. Biol. Chem., February 29, 2008; 283(9): 5719 - 5727. [Abstract] [Full Text] [PDF]