Mitochondrial Biogenesis in Health and Disease

项目摘要 线粒体允许我们的细胞使用氧化磷酸化(OXPHOS)作为一种高效的方式来生成 三磷酸腺苷。内膜包埋的OXPHOS系统酶是由以下组成的多聚体络合物 蛋白质来自两种不同的遗传来源，即核DNA和线粒体DNA。核编码 蛋白质在细胞质核糖体中合成，并输入线粒体。线粒体- 编码的蛋白质，通常是复合体的催化核心亚单位，被合成成不同的线粒体 核糖体。影响这些过程的突变会导致线粒体心脑肌病和 与神经退行性变、代谢性疾病、癌症和衰老有关。 我们已经制定了一项旨在了解分子机制的科学研究计划 这是双重基因起源的线粒体复合体组装的基础。我们的节目包括深入的 研究了三种大分子结构的组装。(I)个别氧磷酸盐的组装 酶，重点是MRC复合体IV或末端的细胞色素c氧化酶(COX)的组装 MRC酶。(2)将MRC酶组装成称为超复合体的超分子结构和 呼吸系统。(3)有丝分裂体的组装和功能。 在每种情况下，我们将追求三个总体目标：(I)定义组装路径/S和顺序 组份和假体基团的结合；(2)确定和表征组装因素 以及(Iii)披露检测到损坏或异常折叠的监管组装检查点 或协调一种成分的合成或成熟，并将其组装成复合体。 这项提案中概述的研究将涉及酵母遗传学，即使用基因编辑技术破坏人类细胞的基因 蛋白质复合体和组装中间体的冷冻-EM重建策略和机制 酵母、人类细胞系、分离的线粒体和纯化的天然和重组蛋白的生物化学 深入了解氧磷脂复合体、超复合体和有丝分裂体组装因子的作用/S。这个 分析生物发生过程的原理和组装因子的活性是至关重要的 这对于我们理解人类线粒体疾病的分子基础具有重要意义。 揭示细胞宏观结构组装发生的机制 标准条件和压力下仍然是现代科学的重大挑战之一，来自生物 从生物医学的角度来看。我们预计我们的计划将继续为缩小与我们之间的差距做出贡献 了解线粒体生物发生在健康和疾病中的作用。

Project Summary Mitochondria allow our cells to use oxidative phosphorylation (OXPHOS) as a highly efficient way to generate ATP. The inner membrane-embedded OXPHOS system enzymes are multimeric complexes composed of proteins from two different genetic origins, namely the nuclear and the mitochondrial DNA. Nucleus-encoded proteins are synthesized in cytoplasmic ribosomes and imported into mitochondria. The mitochondrion- encoded proteins, usually catalytic core subunits of the complexes, are synthesized into distinct mitochondrial ribosomes. Mutations affecting these processes lead to mitochondrial cardio- and encephalo-myopathies and have been linked to neurodegeneration, metabolic disease, cancer, and aging. We have developed a scientific research program aiming to understand the molecular mechanisms underlying the assembly of mitochondrial complexes of dual genetic origin. Our program includes in-depth studies into the assembly of three sets of macromolecular structures. (i) Assembly of individual OXPHOs enzymes, with an emphasis on the assembly of MRC complex IV or cytochrome c oxidase (COX), the terminal MRC enzyme. (ii) Assembly of MRC enzymes into supramolecular structures known as supercomplexes and respirasomes. (iii) Assembly and function of the mitoribosome. In each case, we will pursue three general goals: (i) Define the assembly pathway/s and the order of incorporation of components and prosthetic groups; (ii) Identify and characterize the assembly factors involved, and (iii) Disclose regulatory assembly checkpoints, which detect damaged or abnormally folded components, or coordinate synthesis or maturation of one component with its assembly into the complex. Studies outlined in this proposal will involve yeast genetics, gene disruption in human cells using gene-editing strategies, cryo-EM reconstruction of protein complexes and assembly intermediates, and mechanistic biochemistry in yeast, human cell lines, isolated mitochondria and purified native and recombinant proteins to gain insight into the role/s of OXPHOS complex, supercomplex and mitoribosome assembly factors. The analysis of the principles of the biogenesis process and the activities of the assembly factors is of central importance for our understanding of the molecular basis of human mitochondrial disorders. Uncovering the mechanisms through which assembly of cellular macrostructures takes place in standard conditions and under stress remains one of the grand challenges of modern science from a biological and a biomedical perspective. We anticipate that our program will continue contributing to close the gap in our understanding of mitochondrial biogenesis in health and disease.