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.

项目概要： 线粒体对于能量需求高的细胞至关重要。线粒体 ATP 合酶完成最后的 ATP 产生的步骤。 ATP 合酶突变会导致一系列疾病，包括精神分裂症和 心脏衰竭。因此，ATP 合酶的基础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。独特的挑战 组装 ATP 合酶的过程是协调来自双重遗传起源的基因表达。最近的研究 研究表明，细胞质和线粒体翻译在时间上是同步的，以维持化学计量 ATP 合酶亚基（Couvillion, M.T., et al. Nature, 2016；Soto, I, et al. Genome Biology, 2022）。然而， 跨隔室沟通的基​​本机制仍然很大程度上未知。我的目标是填写 本拟议研究中的知识差距。在酵母中使用单分子荧光原位杂交（smFISH）， 我对核编码亚基 ATP2 和线粒体编码亚基 ATP6/8 的 mRNA 进行了成像。我 发现 ATP2 和 ATP6/8 mRNA 共定位于线粒体网络上。基于此 观察，我假设细胞质和线粒体翻译共定位在 线粒体双膜，从而促进 ATP 合酶的组装。在目标 1 中，我建议 确定 ATP2 mRNA 如何与线粒体共翻译关联。在目标 2 中，我将定义空间 酵母和培养神经元中细胞质和线粒体 mRNA 之间的协调。在目标 3 中，我将 识别调节蛋白并剖析跨区室共定位的潜在机制。 这项研究将揭示细胞在空间上协调核和线粒体的新机制 线粒体生物发生过程中的基因表达。 我接受 Peter Walter 博士的博士学位培训，为我掌握了细胞器生物学、酵母细胞生物学和 生物化学。我与 Robert Singer 博士一起接受的博士后培训使我具备了单分子方面的专业知识 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。期间 K99 阶段，Singer 博士将帮助我培养对酵母和培养神经元中 mRNA 进行成像的技能。延伸 我从酵母到神经元的研究将增加我未来研究的影响。 Michael Rout 博士（联合导师） 将提供鉴定与共定位 mRNA 结合的蛋白质的专业知识。 Liza Pon 博士（联合导师）， Thomas Fox 博士（顾问）和 Christof Osman 博士（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不同方面的知识和技能。 这项研究将为研究线粒体基因表达的空间调控提供机会。它将服务 作为我未来实验室独立研究项目的基础。