Abstract
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)-alpha 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-alpha, 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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing*
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Apoptosis Regulatory Proteins
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Apoptosis* / drug effects
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Blotting, Western
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Calcium-Calmodulin-Dependent Protein Kinases / chemistry*
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Calcium-Calmodulin-Dependent Protein Kinases / genetics
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Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
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Carrier Proteins / genetics
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Carrier Proteins / physiology
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Caspase Inhibitors
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Caspases / genetics
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Caspases / metabolism
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Cell Line
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Death-Associated Protein Kinases
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Fas-Associated Death Domain Protein
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Genes, Dominant / genetics
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Humans
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Inhibitor of Apoptosis Proteins
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Mutation
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Proto-Oncogene Proteins c-bcl-2 / genetics
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Proto-Oncogene Proteins c-bcl-2 / physiology
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RNA, Antisense / genetics
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RNA, Antisense / physiology
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Receptors, Tumor Necrosis Factor / genetics
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Receptors, Tumor Necrosis Factor / physiology
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Serpins / genetics
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Serpins / physiology
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Transfection
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Tumor Cells, Cultured
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Tumor Necrosis Factor-alpha / pharmacology*
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Viral Proteins / genetics
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Viral Proteins / physiology
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fas Receptor / genetics
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fas Receptor / physiology*
Substances
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Adaptor Proteins, Signal Transducing
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Apoptosis Regulatory Proteins
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Carrier Proteins
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Caspase Inhibitors
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FADD protein, human
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Fas-Associated Death Domain Protein
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Inhibitor of Apoptosis Proteins
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Peptide Fragments
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Proto-Oncogene Proteins c-bcl-2
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RNA, Antisense
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Receptors, Tumor Necrosis Factor
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Serpins
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Tumor Necrosis Factor-alpha
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Viral Proteins
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fas Receptor
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inhibitor of apoptosis, Nucleopolyhedrovirus
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interleukin-1beta-converting enzyme inhibitor
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Death-Associated Protein Kinases
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Calcium-Calmodulin-Dependent Protein Kinases
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Caspases