MITOCHONDRIAL MUTAGENESIS & AGE-RELATED PATHOPHYSIOLOGY

线粒体诱变

• 批准号: 2732548
• 负责人: GINO A CORTOPASSI
• 金额: $ 17.3万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-15 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2732548
• 关键词: DNA repair; HeLa cells; aging; animal genetic material tag; antioxidants; apoptosis; calcium flux; cell death; cytochrome c; enzyme activity; gene mutation; genetically modified animals; glutathione peroxidase; human genetic material tag; human tissue; laboratory mouse; microsomes; mitochondrial DNA; superoxide dismutase; superoxides

Specific Aim 1. Mechanism of mitochondrially-triggered cell death. We have recently demonstrated that permeabilized mitochondria release apoptotic factors. providing a testable biochemical, molecular and cellular hypothesis for the mitochondrial regulation of cell death. Firstly, we will test the predictions of our model for cell death by measuring the ability of death-inducers to both induce release of apoptotic factors from mitochondria, and the ability of mitochondrial regulators to inhibit such release. Secondly, our hypothesis for the order of events in cell death will be tested with purified microsomes, mitochondria, and nuclei, and combinations thereof. Thirdly in whole cells, we will observe the sequence of pathological events in toxin- induced apoptosis, and measure the time course and efficacy of mitochondrial effectors to apoptosis. Specific Aim 2. Investigation of the cause-and-effect relationships between mitochondrial antioxidant and repair enzymes and age-related mitochondrial DNA mutagenesis. Using the quantitative assay of mutagenesis developed in the first grant period, we will investigate the effect of specific enzymes to protect cells and animals from mtDNA mutagenesis.

具体目标 1. 线粒体触发的细胞死亡机制。 我们 最近证明，透化线粒体释放 凋亡因素。提供可测试的生化、分子和 线粒体调节细胞死亡的细胞假说。 首先，我们将通过以下方式测试我们的细胞死亡模型的预测： 测量死亡诱导剂诱导释放的能力 来自线粒体的凋亡因子以及线粒体的能力 监管机构抑制这种释放。 其次，我们的假设 细胞死亡的事件顺序将用纯化的微粒体进行测试， 线粒体、细胞核、以及它们的组合。 整体第三 细胞中，我们将观察毒素中病理事件的顺序 诱导细胞凋亡，并测量时程和功效 线粒体效应细胞凋亡。 具体目标2.因果关系调查 线粒体抗氧化剂和修复酶与年龄相关 线粒体 DNA 突变。 使用定量分析 在第一个资助期内开发的诱变，我们将调查 特定酶保护细胞和动物免受 mtDNA 侵害的作用 诱变。