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J. Biol. Chem. 276 (31): 29588-29595

© 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

Identification of the Minimal Tyrosine Residues Required for Linker for Activation of T Cell Function*

Joseph Lin, and Arthur Weiss{ddagger}

From the Departments of Medicine and of Microbiology and Immunology, Biomedical Sciences Graduate Program, Howard Hughes Medical Institute, University of California, San Francisco, California 94143-0795

ABSTRACT Back to Top

Abstract: The linker for activation of T cells (LAT) is essential for signaling through the T cell receptor (TCR). Following TCR stimulation, LAT becomes tyrosine-phosphorylated, creating docking sites for other signaling proteins such as phospholipase C-{gamma}1 (PLC-{gamma}1), Grb2, and Gads. In this study, we have attempted to identify the critical tyrosine residues in LAT that mediate TCR activation-induced mobilization of intracellular Ca2+ and activation of the MAP kinase Erk2. By using the LAT-deficient Jurkat derivative, J.CaM2, stable cell lines were established expressing various tyrosine mutants of LAT. We show that three specific tyrosine residues (Tyr132, Tyr171, and Tyr191) are necessary and sufficient to achieve a Ca2+ flux following TCR stimulation. These tyrosine residues function by reconstituting PLC-{gamma}1phosphorylation and recruitment to LAT. However, these same tyrosines can only partially reconstitute Erk activation. Full reconstitution of Erk requires two additional tyrosine residues (Tyr110 and Tyr226), both of which have the Grb2-binding motif YXN. This reconstitution of Erk activation requires that the critical tyrosine residues be on the same molecule of LAT, suggesting that a single LAT molecule nucleates multiple protein-protein interactions required for optimal signal transduction.

Received for publication March 13, 2001. Revision received May 16, 2001.

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