Kissil J L, Deiss L P, Bayewitch M, Raveh T, Khaspekov G, Kimchi A
Department of Molecular Genetics and Virology, Weizmann Institute of Science, Rehovot, Israel.
J Biol Chem. 1995 Nov 17;270(46):27932-6. 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.
某些细胞因子与其相应的细胞表面受体相互作用会诱导程序性细胞死亡。干扰素-γ在HeLa细胞中诱导一种具有程序性细胞死亡特征的细胞死亡类型。在此，我们报告分离出一个新基因DAP3（死亡相关蛋白3），它参与介导干扰素-γ诱导的细胞死亡。该基因的拯救是通过基于反义cDNA表达文库转染的基因克隆功能选择方法进行的。DAP3基因的反义RNA介导的失活保护细胞免受干扰素-γ诱导的细胞死亡。这一特性使表达该基因的细胞在因持续存在干扰素-γ而具有限制性的环境中具有生长优势，从而为其选择提供了基础。该基因转录成一个单一的1.7千碱基mRNA，在不同组织中普遍表达，并编码一种带有潜在P环基序的46千道尔顿蛋白。DAP3在HeLa细胞中的异位表达与细胞生长不兼容，导致耐药稳定克隆数量减少16倍。所呈现的数据表明DAP3是干扰素-γ诱导的细胞死亡的正向介导因子。
