Requirement for kinase activity of CD4-associated p56lck in antibody-triggered T cell signal transduction.
The lymphoid-specific Src family protein tyrosine kinase p56lck (Lck) is non-covalently associated with the cytoplasmic tail of CD4 and has an essential role in T cell activation. Engagement of ligand by the T cell antigen receptor (TCR) is followed by rapid tyrosine phosphorylation of several cellular proteins, including phospholipase C gamma 1 (PLC) and the TCR-associated CD3 zeta polypeptides. Tyrosine phosphorylation of PLC gamma 1 results in activation of PLC and subsequent phosphatidylinositol turnover. We have studied the effects of the CD4-associated Lck molecule on TCR-mediated activation of the protein tyrosine kinase (PTK) pathway in a murine T cell hybridoma. Antibodies against CD3 elicited the expected PTK activation, which was enhanced upon co-cross-linking of CD4. In contrast, anti-TCR-alpha beta antibodies had no effect on the PTK pathway unless CD4 was co-cross-linked. Antibody cross-linking of CD4 alone failed to induce the same pattern of tyrosine phosphorylation. Similar results were obtained when a chimeric protein consisting of the extracellular and transmembrane domains of CD4 linked to the intracellular Lck molecule was used in place of CD4. The tyrosine kinase activity of Lck was essential for the activity of the chimeric protein. Cross-linking of the CD4/Lck chimera to a CD8/zeta chimeric molecule also facilitated induction of the PTK pathway with anti-CD8 antibodies. Moreover, the interaction of the two chimeric proteins, either in vitro or in vivo, resulted in tyrosine phosphorylation of CD8/zeta. The effects of CD4/Lck on tyrosine phosphorylation and activation of PLC correlated well with the effects on PTK activation. Our results suggest that the Lck molecule positively regulates the TCR-coupled PTK pathway by phosphorylating tyrosines on the TCR-associated CD3 zeta polypeptides.
Chu
K
K
Department of Microbiology and Immunology, University of California, San Francisco 94143-0414.
Littman
D R
DR
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D017252:CD3 Complex / Q000378:metabolism; D015704:CD4 Antigens / Q000378:metabolism*; D016827:CD8 Antigens / Q000378:metabolism; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051379:Mice; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology*; D014443:Tyrosine / Q000378:metabolism; D015854:Up-Regulation
D000906:Antibodies; D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011993:Recombinant Fusion Proteins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)