MOLECULAR MECHANISMS CONTROLING PROGRAMMED CELL DEATH

尽管几乎所有的细胞都注定要死亡，但我们对此知之甚少。 关于衰老和死亡的分子机制。 这项提案概述了表征基因的实验，这些基因 调节发育过程中的细胞死亡。这些实验源于我们的 烟草天蛾节间肌群的研究 Manduca sexta，它在36小时内死亡，以响应 内分泌触发。我们已经分离了4个基因的cdna克隆，它们是 在ISM变得决心要死的时候表达出来，以及三个 被压抑。 将使用一系列分子方法来分离、测序 并表达额外的细胞死亡基因。特别是，我们正在 对转录调控因子的特征感兴趣 细胞死亡的过程。昆虫肌肉纤维将被注射一种 从我们的克隆产生的以下分子：反义 转录本、正义转录本、抗体、纯化的表达蛋白 或与假定的细胞死亡融合的报告基因的重组体 基因启动子。我们将利用果蝇进行基因改造 对细胞死亡过程的分析。从昆虫中分离出的分子 将被用来研究脊椎动物神经发生过程中的细胞死亡。它是 显然，对这种模拟系统审查将提供基本的 洞察调节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.