p56Lck tyrosine kinase enhances the assembly of death-inducing signaling complex during fas-mediated apoptosis

Abstract

Although the death-inducing signaling complex (DISC) is rapidly assembled, several lines of evidence suggest that formation of this complex is not the first consequence of cell surface CD95 (Fas) stimulation but rather a later step in this process. Activation of Fas triggers a cascade of signaling events that culminate in cellular apoptosis. Tyrosine kinases are critical effectors in T cell activation. However, their functional involvement in death receptor-mediated apoptosis is unknown. Here, we used p56Lck- deficient cells to show that CD95-induced cell death is highly dependent on p56Lck activity and its localization within plasma membrane. We found that p56Lck acts upstream of the mitochondria; in the absence of p56Lck, Bid cleavage and the release of cytochrome c were severely impaired. Moreover, p56Lck-deficient cells or cells expressing an inactive form of p56Lck displayed defective formation of the DISC post CD95 stimulation. In vivo reconstitution of thymocytes from p 56lck-deficient mice, which are resistant to apoptosis, with p56 Lck restored Fas-mediated cell death. Our results support a novel model whereby sensitivity to apoptosis is regulated through quantitative changes in the stoichiometry of DISC components triggered by p56Lck activation and localization. © 2007 by The American Society for Biochemistry and Molecular Biology, Inc.