Original Article

Recruitment of SLP-76 to the Membrane and Glycolipid-enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling

Correspondence to: Gary A. Koretzky, Dept. of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6160. Tel:215-746-5522 Fax:215-746-5525

Two hematopoietic-specific adapters, src homology 2 domain–containing leukocyte phosphoprotein of 76 kD (SLP-76) and linker for activation of T cells (LAT), are critical for T cell development and T cell receptor (TCR) signaling. Several studies have suggested that SLP-76 and LAT function coordinately to promote downstream signaling. In support of this hypothesis, we find that a fraction of SLP-76 localizes to glycolipid-enriched membrane microdomains (GEMs) after TCR stimulation. This recruitment of SLP-76 requires amino acids 224–244. The functional consequences of targeting SLP-76 to GEMs for TCR signaling are demonstrated using a LAT/SLP-76 chimeric protein. Expression of this construct reconstitutes TCR-inducted phospholipase C1 phosphorylation, extracellular signal–regulated kinase activation, and nuclear factor of activated T cells (NFAT) promoter activity in LAT-deficient Jurkat T cells (J.CaM2). Mutation of the chimeric construct precluding its recruitment to GEMs diminishes but does not eliminate its ability to support TCR signaling. Expression of a chimera that lacks SLP-76 amino acids 224–244 restores NFAT promoter activity, suggesting that if localized, SLP-76 does not require an association with Gads to promote T cell activation. In contrast, mutation of the protein tyrosine kinase phosphorylation sites of SLP-76 in the context of the LAT/SLP-76 chimera abolishes reconstitution of TCR function. Collectively, these experiments show that optimal TCR signaling relies on the compartmentalization of SLP-76 and that one critical function of LAT is to bring SLP-76 and its associated proteins to the membrane.

Key Words: adapter proteins, signal transduction, T cell activation, lipid rafts, protein tyrosine kinase

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Sankarshanan, M., Ma, Z., Iype, T., Lorenz, U. (2007). Identification of a Novel Lipid Raft-Targeting Motif in Src Homology 2-Containing Phosphatase 1. J. Immunol. 179: 483-490 [Abstract] [Full Text]  
• Beach, D., Gonen, R., Bogin, Y., Reischl, I. G., Yablonski, D. (2007). Dual Role of SLP-76 in Mediating T Cell Receptor-induced Activation of Phospholipase C-{gamma}1. J. Biol. Chem. 282: 2937-2946 [Abstract] [Full Text]  
• Kliche, S., Breitling, D., Togni, M., Pusch, R., Heuer, K., Wang, X., Freund, C., Kasirer-Friede, A., Menasche, G., Koretzky, G. A., Schraven, B. (2006). The ADAP/SKAP55 Signaling Module Regulates T-Cell Receptor-Mediated Integrin Activation through Plasma Membrane Targeting of Rap1.. Mol. Cell. Biol. 26: 7130-7144 [Abstract] [Full Text]  
• Bunnell, S. C., Singer, A. L., Hong, D. I., Jacque, B. H., Jordan, M. S., Seminario, M.-C., Barr, V. A., Koretzky, G. A., Samelson, L. E. (2006). Persistence of cooperatively stabilized signaling clusters drives T-cell activation.. Mol. Cell. Biol. 26: 7155-7166 [Abstract] [Full Text]  
• Abtahian, F., Bezman, N., Clemens, R., Sebzda, E., Cheng, L., Shattil, S. J., Kahn, M. L., Koretzky, G. A. (2006). Evidence for the Requirement of ITAM Domains but Not SLP-76/Gads Interaction for Integrin Signaling in Hematopoietic Cells.. Mol. Cell. Biol. 26: 6936-6949 [Abstract] [Full Text]  
• Watanabe, R., Harada, Y., Takeda, K., Takahashi, J., Ohnuki, K., Ogawa, S., Ohgai, D., Kaibara, N., Koiwai, O., Tanabe, K., Toma, H., Sugamura, K., Abe, R. (2006). Grb2 and Gads Exhibit Different Interactions with CD28 and Play Distinct Roles in CD28-Mediated Costimulation. J. Immunol. 177: 1085-1091 [Abstract] [Full Text]  
• Silverman, M. A., Shoag, J., Wu, J., Koretzky, G. A. (2006). Disruption of SLP-76 Interaction with Gads Inhibits Dynamic Clustering of SLP-76 and Fc{varepsilon}RI Signaling in Mast Cells.. Mol. Cell. Biol. 26: 1826-1838 [Abstract] [Full Text]  
• Ziemba, S. E., Mattingly, R. R., McCabe, M. J. Jr., Rosenspire, A. J. (2006). Inorganic Mercury Inhibits the Activation of LAT in T-Cell Receptor-Mediated Signal Transduction. Toxicol Sci 89: 145-153 [Abstract] [Full Text]  
• Gonen, R., Beach, D., Ainey, C., Yablonski, D. (2005). T Cell Receptor-induced Activation of Phospholipase C-{gamma}1 Depends on a Sequence-independent Function of the P-I Region of SLP-76. J. Biol. Chem. 280: 8364-8370 [Abstract] [Full Text]  
• Fawcett, V. C. J., Lorenz, U. (2005). Localization of Src Homology 2 Domain-Containing Phosphatase 1 (SHP-1) to Lipid Rafts in T Lymphocytes: Functional Implications and a Role for the SHP-1 Carboxyl Terminus. J. Immunol. 174: 2849-2859 [Abstract] [Full Text]  
• Singer, A. L., Bunnell, S. C., Obstfeld, A. E., Jordan, M. S., Wu, J. N., Myung, P. S., Samelson, L. E., Koretzky, G. A. (2004). Roles of the Proline-rich Domain in SLP-76 Subcellular Localization and T Cell Function. J. Biol. Chem. 279: 15481-15490 [Abstract] [Full Text]  
• Alexander, M., Bor, Y.-c., Ravichandran, K. S., Hammarskjold, M.-L., Rekosh, D. (2004). Human Immunodeficiency Virus Type 1 Nef Associates with Lipid Rafts To Downmodulate Cell Surface CD4 and Class I Major Histocompatibility Complex Expression and To Increase Viral Infectivity. J. Virol. 78: 1685-1696 [Abstract] [Full Text]  
• Stebbins, C. C., Watzl, C., Billadeau, D. D., Leibson, P. J., Burshtyn, D. N., Long, E. O. (2003). Vav1 Dephosphorylation by the Tyrosine Phosphatase SHP-1 as a Mechanism for Inhibition of Cellular Cytotoxicity. Mol. Cell. Biol. 23: 6291-6299 [Abstract] [Full Text]  
• Kuhne, M. R., Lin, J., Yablonski, D., Mollenauer, M. N., Ehrlich, L. I. R., Huppa, J., Davis, M. M., Weiss, A. (2003). Linker for Activation of T Cells, {zeta}-Associated Protein-70, and Src Homology 2 Domain-Containing Leukocyte Protein-76 are Required for TCR-Induced Microtubule-Organizing Center Polarization. J. Immunol. 171: 860-866 [Abstract] [Full Text]  
• Kettner, A., Pivniouk, V., Kumar, L., Falet, H., Lee, J.-S., Mulligan, R., Geha, R. S. (2003). Structural Requirements of SLP-76 in Signaling via the High-Affinity Immunoglobulin E Receptor (Fc{varepsilon}RI) in Mast Cells. Mol. Cell. Biol. 23: 2395-2406 [Abstract] [Full Text]  
• Yamasaki, S., Nishida, K., Sakuma, M., Berry, D., McGlade, C. J., Hirano, T., Saito, T. (2003). Gads/Grb2-Mediated Association with LAT Is Critical for the Inhibitory Function of Gab2 in T Cells. Mol. Cell. Biol. 23: 2515-2529 [Abstract] [Full Text]  
• Fragoso, R., Ren, D., Zhang, X., Su, M. W.-C., Burakoff, S. J., Jin, Y.-J. (2003). Lipid Raft Distribution of CD4 Depends on its Palmitoylation and Association with Lck, and Evidence for CD4-Induced Lipid Raft Aggregation as an Additional Mechanism to Enhance CD3 Signaling. J. Immunol. 170: 913-921 [Abstract] [Full Text]  
• Tanimura, N., Nagafuku, M., Minaki, Y., Umeda, Y., Hayashi, F., Sakakura, J., Kato, A., Liddicoat, D. R., Ogata, M., Hamaoka, T., Kosugi, A. (2003). Dynamic changes in the mobility of LAT in aggregated lipid rafts upon T cell activation. J. Cell Biol. 160: 125-135 [Abstract] [Full Text]  
• Zhu, M., Janssen, E., Zhang, W. (2003). Minimal Requirement of Tyrosine Residues of Linker for Activation of T Cells in TCR Signaling and Thymocyte Development. J. Immunol. 170: 325-333 [Abstract] [Full Text]  
• Zhu, M., Janssen, E., Leung, K., Zhang, W. (2002). Molecular Cloning of a Novel Gene Encoding a Membrane-associated Adaptor Protein (LAX) in Lymphocyte Signaling. J. Biol. Chem. 277: 46151-46158 [Abstract] [Full Text]  
• Kovacs, B., Maus, M. V., Riley, J. L., Derimanov, G. S., Koretzky, G. A., June, C. H., Finkel, T. H. (2002). Human CD8+ T cells do not require the polarization of lipid rafts for activation and proliferation. Proc. Natl. Acad. Sci. USA 99: 15006-15011 [Abstract] [Full Text]  
• Judd, B. A., Myung, P. S., Obergfell, A., Myers, E. E., Cheng, A. M., Watson, S. P., Pear, W. S., Allman, D., Shattil, S. J., Koretzky, G. A. (2002). Differential Requirement for LAT and SLP-76 in GPVI versus T Cell Receptor Signaling. J. Exp. Med. 195: 705-717 [Abstract] [Full Text]  
• Hussain, S. F., Anderson, C. F., Farber, D. L. (2002). Differential SLP-76 Expression and TCR-Mediated Signaling in Effector and Memory CD4 T Cells. J. Immunol. 168: 1557-1565 [Abstract] [Full Text]  
• Teixeiro, E., Fuentes, P., Galocha, B., Alarcon, B., Bragado, R. (2002). T Cell Receptor-mediated Signal Transduction Controlled by the beta Chain Transmembrane Domain. APOPTOSIS-DEFICIENT CELLS DISPLAY UNBALANCED MITOGEN-ACTIVATED PROTEIN KINASES ACTIVITIES UPON T CELL RECEPTOR ENGAGEMENT. J. Biol. Chem. 277: 3993-4002 [Abstract] [Full Text]  
• Dykstra, M., Cherukuri, A., Pierce, S. K. (2001). Rafts and synapses in the spatial organization of immune cell signaling receptors. J. Leukoc. Biol. 70: 699-707 [Abstract] [Full Text]  
• Veri, M.-C., DeBell, K. E., Seminario, M.-C., DiBaldassarre, A., Reischl, I., Rawat, R., Graham, L., Noviello, C., Rellahan, B. L., Miscia, S., Wange, R. L., Bonvini, E. (2001). Membrane Raft-Dependent Regulation of Phospholipase C{gamma}-1 Activation in T Lymphocytes. Mol. Cell. Biol. 21: 6939-6950 [Abstract] [Full Text]  
• Vyas, Y. M., Mehta, K. M., Morgan, M., Maniar, H., Butros, L., Jung, S., Burkhardt, J. K., Dupont, B. (2001). Spatial Organization of Signal Transduction Molecules in the NK Cell Immune Synapses During MHC Class I-Regulated Noncytolytic and Cytolytic Interactions. J. Immunol. 167: 4358-4367 [Abstract] [Full Text]  
• Yankee, T. M., Draves, K. E., Ewings, M. K., Clark, E. A., Graves, J. D. (2001). CD95/Fas induces cleavage of the GrpL/Gads adaptor and desensitization of antigen receptor signaling. Proc. Natl. Acad. Sci. USA 98: 6789-6793 [Abstract] [Full Text]  
• Woods, M. L., Shimizu, Y. (2001). Signaling networks regulating {beta}1 integrin-mediated adhesion of T lymphocytes to extracellular matrix. J. Leukoc. Biol. 69: 874-880 [Abstract] [Full Text]  
• Kikuchi, K., Kawasaki, Y., Ishii, N., Sasaki, Y., Asao, H., Takeshita, T., Miyoshi, I., Kasai, N., Sugamura, K. (2001). Suppression of thymic development by the dominant-negative form of Gads. Int Immunol 13: 777-783 [Abstract] [Full Text]  
• Lin, J., Weiss, A. (2001). Identification of the Minimal Tyrosine Residues Required for Linker for Activation of T Cell Function. J. Biol. Chem. 276: 29588-29595 [Abstract] [Full Text]  
• Goitsuka, R., Tatsuno, A., Ishiai, M., Kurosaki, T., Kitamura, D. (2001). MIST Functions through Distinct Domains in Immunoreceptor Signaling in the Presence and Absence of LAT. J. Biol. Chem. 276: 36043-36050 [Abstract] [Full Text]  
• Sauer, K., Liou, J., Singh, S. B., Yablonski, D., Weiss, A., Perlmutter, R. M. (2001). Hematopoietic Progenitor Kinase 1 Associates Physically and Functionally with the Adaptor Proteins B Cell Linker Protein and SLP-76 in Lymphocytes. J. Biol. Chem. 276: 45207-45216 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search

TABLE OF CONTENTS