MOLECULAR MECHANISMS CONTROLING PROGRAMMED CELL DEATH

虽然几乎所有的细胞都注定要死亡， 衰老和死亡的分子机制 该提案概述了描述基因特征的实验， 在发育过程中介导细胞死亡。 这些实验源于我们的 烟草天蛾节间肌的研究 曼陀罗，在36小时内死亡，以响应一个定义的 内分泌触发器 我们分离了4个基因的cDNA克隆， 当ISM变得致力于死亡时， 压抑 一系列的分子方法将被用来分离，测序 并表达额外的细胞死亡基因。 我们尤其 感兴趣的是表征转录调节因子， 细胞死亡的过程。 昆虫的肌肉纤维将被注入一个 从我们的克隆中产生的以下分子： 转录物、有义转录物、抗体、纯化的表达蛋白 或用报告基因与假定的细胞死亡融合的重组体 基因启动子 我们将利用果蝇进行遗传 分析细胞死亡过程。 从昆虫中分离的分子 将被用于研究脊椎动物神经发生过程中的细胞死亡。 是 显然，对这种模式系统研究将提供基本的 深入了解介导衰老和死亡的分子机制 在动物细胞中。

While virtually all cells are fated to die, very little is known about the molecular mechanisms responsible for senescence and death. This proposal outlines experiments to characterize the genes which mediate cell death during development. These experiments arise from our studies of-the intersegmental muscles (ISM) of the tobacco hawkmoth Manduca sexta, which die during a 36 hour period in response to a defined endocrine trigger. We have isolated cDNA clones for 4 genes that-are expressed when the ISM become committed to die, and three that are repressed. A range of molecular approaches will be used to isolate, sequence and express additional cell death genes. In particular, we are interested in characterizing transcriptional regulators which initiate the cell death process. Insect muscle fibers will be injected with one of the following molecules generated from our clones: anti-sense transcripts, sense transcripts, antibodies, purified expressed proteins or recombinants made with a reporter gene fused to putative cell-death gene promoters. We will utilize the fly Drosophila for a genetic analysis of the cell death process. Molecules isolated from -insects will be used to examine cell death during vertebrate neurogenesis. It is clear that that examination of such model systems will provide basic insight into the molecular mechanisms which mediate senescence and death in animal cells.