Isolation of DAP3, a novel mediator of interferon-gamma-induced cell death

• Published in: The Journal of biological chemistry Vol. 270; no. 46; pp. 27932 - 27936
• Main Authors: Kissil, J L, Deiss, L P, Bayewitch, M, Raveh, T, Khaspekov, G, Kimchi, A
• Format: Journal Article
• Language: English
• Published: United States 17.11.1995
• Subjects: Amino Acid Sequence; Animals; Antibodies; Apoptosis Regulatory Proteins; Base Sequence; Cell Death - drug effects; Cell Death - physiology; Cell Division; Cloning, Molecular; DNA, Antisense; DNA, Complementary; Gene Expression; Gene Library; HeLa Cells; Humans; Interferon-gamma - pharmacology; Molecular Sequence Data; Protein Biosynthesis; Proteins - genetics; Proteins - isolation & purification; Rabbits - immunology; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Reticulocytes - metabolism; Ribosomal Proteins; RNA, Antisense - metabolism; RNA, Messenger - biosynthesis; RNA-Binding Proteins; Transfection
• ISSN: 0021-9258
• DOI: 10.1074/jbc.270.46.27932

Interaction of certain cytokines with their corresponding cell-surface receptors induces programmed cell death. Interferon-gamma induces in HeLa cells a type of cell death with features characteristic of programmed cell death. Here, we report the isolation of a novel gene, DAP3 (death-associated protein-3), involved in mediating interferon-gamma-induced cell death. The rescue of this gene was performed by a functional selection approach of gene cloning that is based on transfection with an antisense cDNA expression library. The antisense RNA-mediated inactivation of the DAP3 gene protected the cells from interferon-gamma-induced cell death. This property endowed the cells expressing it with a growth advantage in an environment restrictive due to the continuous presence of interferon-gamma and thus provided the basis of its selection. The gene is transcribed into a single 1.7-kilobase mRNA, which is ubiquitously expressed in different tissues and codes for a 46-kDa protein carrying a potential P-loop motif. Ectopic expression of DAP3 in HeLa cells was not compatible with cell growth, resulting in a 16-fold reduction in the number of drug-resistant stable clones. The data presented suggest that DAP3 is a positive mediator of cell death induced by interferon-gamma.