Spatial coordination of cytosolic and mitochondrial translation

细胞质和线粒体翻译的空间协调

• 批准号: 10739786
• 负责人: Weihan Li
• 金额: $ 12.5万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739786
• 关键词: Adult; Alzheimer' s Disease; Binding Proteins; Biochemistry; Biogenesis; Biology; Brain; Cardiac Myocytes; Cells; Cellular biology; Communication; Cytosol; Disease; Doctor of Philosophy; Endoplasmic Reticulum; Face; Fluorescent in Situ Hybridization; Foundations; Future; Gene Expression; Genetic; Genome; Health; Heart failure; Human; Image; Immobilization; In Situ; In Situ Hybridization; Knowledge; Knowledge acquisition; Laboratories; Membrane; Mentors; Messenger RNA; Mitochondria; Mitochondrial Diseases; Mitochondrial Matrix; Mitochondrial Proton-Translocating ATPases; Molecular; Mutation; Nature; Neurons; Nuclear; Organelles; Oxidative Phosphorylation; Phase; Production; Proteins; RNA-Binding Proteins; Regulation; Research; Schizophrenia; Series; Side; Site; Synapses; Technology; Testing; Training; Translating; Translation Initiation; Translational Repression; Translations; Weight; Yeasts; experimental study; insight; live cell imaging; mRNA Translation; mitochondrial genome; mitochondrial messenger RNA; molecular imaging; novel; post-doctoral training; programs; single molecule; skills; stoichiometry

Project Summary: Mitochondria are critical for cells with high energy demand. Mitochondrial ATP synthase completes the final step of ATP production. Mutations in ATP synthase cause a range of diseases, including Schizophrenia, and heart failure. Thus, the basic biology of ATP synthase is important for health and disease. The subunits of the ATP synthase are encoded by the nuclear and mitochondrial genome. A unique challenge of assembling the ATP synthase is to coordinate the gene expressions from dual genetic origins. Recent studies showed that cytosolic and mitochondrial translation are temporally synchronized to maintain the stoichiometry of the ATP synthase subunits (Couvillion, M.T., et al. Nature, 2016; Soto, I, et al. Genome Biology, 2022). However, the underlying mechanism of the cross-compartment communication remains largely unknown. I aim to fill in the knowledge gap in this proposed study. Using single-molecule fluorescent in situ hybridization (smFISH) in yeast, I imaged the mRNAs of a nuclear-encoded subunit, ATP2, and a mitochondrial-encoded subunit, ATP6/8. I discovered that the ATP2 and ATP6/8 mRNA co-localized on the mitochondrial network. Based on this observation, I hypothesize that the cytosolic and mitochondrial translation co-localize on opposite sides of the mitochondrial double membrane, thereby promoting the assembly of the ATP synthase. In Aim 1, I propose to determine how the ATP2 mRNAs co-translationally associate with mitochondria. In Aim 2, I will define the spatial coordination between the cytosolic and mitochondrial mRNAs in yeast and cultured neurons. In Aim 3, I shall identify the regulating proteins and dissect the underlying mechanism of the cross-compartment co-localization. This study will uncover a novel mechanism by which cells spatially coordinate the nuclear and mitochondrial gene expressions during mitochondrial biogenesis. My PhD training with Dr. Peter Walter prepared me with the skills in organelle biology, yeast cell biology, and biochemistry. My postdoctoral training with Dr. Robert Singer equipped me with the expertise in single-molecule imaging. These complementary skills give me the unique opportunity to conduct this proposed study. During the K99 phase, Dr. Singer will help me develop the skills to image mRNAs in yeast and cultured neurons. Extending my research from yeast to neurons will increase the impact of my future research. Dr. Michael Rout (Co-Mentor) will provide the expertise to identify the proteins that bind to the co-localizing mRNAs. Dr. Liza Pon (Co-Mentor), Dr. Thomas Fox (Consultant), and Dr. Christof Osman (Collaborator) form my mitochondrial mentoring team. Their diverse background will allow me to acquire the knowledge and skills of different aspects of mitochondria. This study will open opportunities to study the spatial regulation of mitochondrial gene expression. It will serve as a foundation for an independent research program in my future laboratory.

项目摘要： 线粒体对于高能量需求的细胞至关重要。线粒体ATP合酶完成了最终 ATP生产的步骤。 ATP合酶中的突变会引起一系列疾病，包括精神分裂症和 心脏衰竭。因此，ATP合酶的基本生物学对于健康和疾病很重要。 ATP合酶的亚基由核和线粒体基因组编码。一个独特的挑战 组装ATP合酶是为了协调双重遗传起源的基因表达。最近的研究 表明胞质和线粒体翻译在时间上同步以维持化学计量法 ATP合酶亚基（Couvillion，M.T。等人，2016年； Soto，I等，基因组生物学，2022年）。然而， 跨隔间通信的基本机制在很大程度上仍然未知。我的目标是填写 在这项拟议的研究中，知识差距。使用酵母中的单分子荧光原位杂交（Smfish）， 我成像了核编码亚基ATP2和线粒体编码的亚基ATP6/8的mRNA。我 发现在线粒体网络上共定位了ATP2和ATP6/8 mRNA。基于此 观察到，我假设胞质和线粒体翻译在相对侧共定位 线粒体双膜，从而促进ATP合酶的组装。在AIM 1中，我建议 确定ATP2 mRNA如何共译与线粒体缔合。在AIM 2中，我将定义空间 酵母和培养神经元中的胞质和线粒体mRNA之间的协调。在AIM 3中，我将 确定调节蛋白质并剖析跨隔间共定位的潜在机制。 这项研究将发现一种新的机制，细胞通过空间协调核和线粒体 线粒体生物发生过程中的基因表达。 我与彼得·沃尔特（Peter Walter）博士的博士培训为我准备了Organelle Biology，酵母细胞生物学和 生物化学。我与罗伯特·辛格（Robert Singer）博士的博士后培训为我配备了单分子的专业知识 成像。这些互补的技能为我提供了进行这项拟议研究的独特机会。在 K99阶段，Singer博士将帮助我发展技能，以酵母和培养的神经元形象形象。扩展 我从酵母到神经元的研究将增加我未来研究的影响。迈克尔·鲁特（Michael Rout）博士（院） 将提供专业知识，以识别与共定位的mRNA结合的蛋白质。 Liza Pon博士（Co-entor）， 托马斯·福克斯（Thomas Fox）博士（顾问）和克里斯托夫·奥斯曼（Christof Osman）（合作者）组成了我的线粒体指导团队。 他们的不同背景将使我能够获得线粒体各个方面的知识和技能。 这项研究将开放研究线粒体基因表达的空间调节的机会。它将服务 作为我未来实验室中独立研究计划的基础。