Systems Analysis of Mammalian Mitochondrial DNA Segregation

 描述（由申请人提供）：线粒体DNA（mtDNA）的忠实遗传对于细胞呼吸是必不可少的，因此对于大多数真核生物来说都是如此。然而，在哺乳动物细胞中控制mtDNA维持和分离的途径还没有很好地理解。这项工作的目标是了解哺乳动物mtDNA调控的分子机制。一旦知道这些信息，就有可能确定影响人类健康的mtDNA单倍型分离的因素。具体来说，该提案将解决线粒体DNA-蛋白质复合物（称为类核）在哺乳动物细胞中的行为。在目标1中，类核复合物的超分辨率和共聚焦显微镜加上遗传方法来消耗和修改复杂的组件将被用来调查细胞内的类核分离的基础和测试候选效应蛋白的参与。目标2将阐述类核是如何组织或整合细胞信号通路的。这将通过系统地绘制哺乳动物mtDNA类核复合物的物理和遗传相互作用来实现。这些实验将提供对mtDNA遗传单位的物理和功能组织的深入了解，并有效地绘制类核结构的整合“相互作用组”。

 DESCRIPTION (provided by applicant): Faithful inheritance of mitochondrial DNA (mtDNA) is essential for cellular respiration and thus for most eukaryotic life. However, pathways that control mtDNA maintenance and segregation in mammalian cells are not well understood. The goal of this work is to understand the molecular mechanisms underlying the regulation of mammalian mtDNA. Once this information is known, it will be possible to identify factors that contribute to the segregation of mtDNA haplotypes that impact human health. Specifically, this proposal will address how mtDNA-protein complexes, termed nucleoids, behave in mammalian cells. In Aim 1, super-resolution and confocal microscopy of nucleoid complexes coupled with genetic approaches to deplete and modify complex components will be employed to investigate the intracellular basis of nucleoid segregation and test the involvement of candidate effector proteins. Aim 2 will address how the nucleoid is organized or integrated with cellular signaling pathways. This will be accomplished by systematically mapping physical and genetic interactions of the mammalian mtDNA nucleoid complex. These experiments will provide insight into the physical and functional organization of the heritable unit of mtDNA and effectively map the integrated 'interactome' of the nucleoid structure.