Defining intracellular mitochondrial heterogeneity establishment and maintenance

项目摘要 线粒体是负责大多数细胞代谢事件的亚细胞室，包括 氧化磷酸化、血红素生物合成和其他代谢通量。鉴于常见的对 线粒体生理学，令人惊讶的是，最近的两项研究发现了线粒体的亚群 专门研究它们的职能和组成(2，10)。根据这些发现，我们假设创新的 细胞内线粒体异质性存在于细胞内并通过细胞内维持的假设 细胞器--交流。在这个提案中，我们将定义细胞内线粒体的异质性，并发现 特化线粒体的特征。使用独立的技术：1)CRISPR介导的内切 精选线粒体蛋白的标记；2)分离的线粒体的邻近标记和亲和纯化 线粒体群体将与Steven Gygi进行比较蛋白质组学和代谢组学(见 附信)。在细胞中建立线粒体异质性的一个潜在机制是有明显的 细胞内通讯的模式。在此背景下，我们提出细胞器邻近(内质网，高尔基体， 过氧化物酶、溶酶体、内切体和脂滴)是建立和维持 线粒体通过改变各自的蛋白质组来特化线粒体，从而改变线粒体的功能和 新陈代谢。此外，我们的目标是揭示细胞内线粒体异质性在细胞内的作用。 动态平衡。总体而言，从拟议实验中获得的见解将提供一个创新和 关于线粒体生物学和动态平衡的激动人心的观点。

Project Summary Mitochondria are the subcellular compartment responsible for a majority of cellular metabolic events including oxidative phosphorylation, heme biosynthesis, and other metabolic fluxes. Given the common depictions of mitochondrial physiology, it is striking that two recent studies have discovered subsets of mitochondria that are specialized in their functions and composition (2,10). In accordance with these findings, we posit the innovative hypothesis that intracellular mitochondrial heterogeneity exists in cells and is maintained through intracellular organelle-communication. In this proposal, we will define intracellular mitochondrial heterogeneity and discover the characteristics of specialized mitochondria. Using independent techniques: 1) CRISPR-mediated endo- tagging of select mitochondrial protein; 2) Proximity labeling and affinity purification of mitochondria, isolated mitochondrial populations will undergo comparative proteomics and metabolomics with Steven Gygi (see attached letter). A potential mechanism to establish mitochondrial heterogeneity in cells is to have distinct modes of intracellular communication. In this context, we propose that organelle proximity (ER, Golgi, peroxisome, lysosome, endosome, and lipid droplets) is necessary for the establishment and maintenance of specialized mitochondria by changing their individual proteomes and thus, mitochondrial function and metabolism. Additionally, we aim to uncover the roles for intracellular mitochondrial heterogeneity in cellular homeostasis. Overall, the insights gained from the proposed experiments will provide an innovative and exciting perspective on mitochondrial biology and homeostasis.