DAP-Kinase Participates in TNF-α- and Fas-Induced Apoptosis and Its Function Requires the Death Domain

• Published in: The Journal of cell biology Vol. 146; no. 1; pp. 141 - 148
• Main Authors: Cohen, Ofer, Inbal, Boaz, Kissil, Joseph L., Raveh, Tal, Berissi, Hanna, Spivak-Kroizaman, Taly, Feinstein, Elena, Kimchi, Adi
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 12.07.1999; The Rockefeller University Press
• Subjects: Adaptor Proteins, Signal Transducing; Antibodies; Apoptosis; Apoptosis - drug effects; Apoptosis Regulatory Proteins; Biochemistry; Blotting, Western; Calcium-Calmodulin-Dependent Protein Kinases - chemistry; Calcium-Calmodulin-Dependent Protein Kinases - genetics; Calcium-Calmodulin-Dependent Protein Kinases - metabolism; Carrier Proteins - genetics; Carrier Proteins - physiology; Caspase Inhibitors; Caspases - genetics; Caspases - metabolism; Cell death; Cell Line; Cell lines; Cellular biology; Death-Associated Protein Kinases; fas Receptor - genetics; fas Receptor - physiology; Fas-Associated Death Domain Protein; Genes, Dominant - genetics; Genetics; HEK293 cells; HeLa cells; Humans; Inhibitor of Apoptosis Proteins; Kidney cells; Kimchi; Mutation; Original; Peptide Fragments - chemistry; Peptide Fragments - genetics; Peptide Fragments - metabolism; Proteins; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - physiology; Receptors; Receptors, Tumor Necrosis Factor - genetics; Receptors, Tumor Necrosis Factor - physiology; RNA, Antisense - genetics; RNA, Antisense - physiology; Serpins - genetics; Serpins - physiology; Transfection; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha - pharmacology; Viral Proteins - genetics; Viral Proteins - physiology
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.146.1.141

Death-associated protein (DAP)-kinase is a calcium/calmodulin regulated serine/threonine kinase that carries ankyrin repeats, a death domain, and is localized to the cytoskeleton. Here, we report that this kinase is involved in tumor necrosis factor (TNF)-α and Fas-induced apoptosis. Expression of DAP-kinase antisense RNA protected cells from killing by anti-Fas/APO-1 agonistic antibodies. Deletion of the death domain abrogated the apoptotic functions of the kinase, thus, documenting for the first time the importance of this protein domain. Overexpression of a fragment encompassing the death domain of DAP-kinase acted as a specific dominant negative mutant that protected cells from TNF-α, Fas, and FADD/MORT1-induced cell death. DAP-kinase apoptotic function was blocked by bcl-2 as well as by crmA and p35 inhibitors of caspases, but not by the dominant negative mutants of FADD/MORT1 or of caspase 8. Thus, it functions downstream to the receptor complex and upstream to other caspases. The multidomain structure of this serine/threonine kinase, combined with its involvement in cell death induced by several different triggers, place DAP-kinase at one of the central molecular pathways leading to apoptosis.