Isolation of DAP3, a novel mediator of interferon-gamma-induced cell death.
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.
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D016923:Cell Death / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division; D003001:Cloning, Molecular; D016373:DNA, Antisense; D018076:DNA, Complementary; D015870:Gene Expression; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification; D016372:RNA, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D016601:RNA-Binding Proteins; D011817:Rabbits / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012156:Reticulocytes / Q000378:metabolism; D012269:Ribosomal Proteins; D014162:Transfection
D000906:Antibodies; D051017:Apoptosis Regulatory Proteins; C097041:DAP3 protein, human; D016373:DNA, Antisense; D018076:DNA, Complementary; D011506:Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; D007371:Interferon-gamma