Elucidating the regulation and function of amyloid-like assemblies

阐明淀粉样蛋白组装体的调节和功能

• 批准号: 10405902
• 负责人: Luke E Berchowitz
• 金额: $ 44.95万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10405902
• 关键词: Aging; Alzheimer' s Disease; Amyloid; Award; Biochemical; Cells; Deposition; Disease; Etiology; Fluorescence; Freedom; Genetic Screening; Germ Cells; Goals; Institutes; Knowledge; Lead; Mediating; Meiosis; Messenger RNA; Molecular; Neurodegenerative Disorders; Parkinson Disease; Pathologic; Pathway interactions; Phase; Phosphorylation Site; Physiological; Play; Population; Prealbumin; Preventive therapy; Process; Property; Proteins; RNA-Binding Proteins; Regulation; Reporter; Research; Research Personnel; Retrotransposon; Role; Saccharomycetales; Signal Pathway; Structure; System; Therapeutic; Time; Translations; Universities; Yeasts; base; cardiac amyloidosis; fibrous protein; genome integrity; human disease; insight; monomer; programs; research vision; tool

PROJECT SUMMARY Although amyloids are best understood for their roles in the etiology of numerous human diseases as pathological protein deposits, amyloid and amyloid-like assemblies can serve critical functions1. To mediate these functions, some cells have the ability to regulate assembly and clearance of these structures2,3. My research vision is to delineate the physiological pathways and mechanisms that healthy cells have evolved to regulate formation, function, and reversibility of amyloids. These studies are motivated by our discovery that in order to control translation during meiosis, budding yeast regulates assembly of the RNA-binding protein Rim4 into a translational repressor that has several biochemical properties of an amyloid (thus termed amyloid-like)2. Knowledge gained from these studies will lead to important advances in our understanding of the causes of neurodegenerative diseases and in time could lead to therapeutic opportunities. Based on our recent progress studying Rim4 clearance, we now appreciate that yeast cells use multi- site phosphorylation to abruptly reverse amyloid-like Rim4 into oligomers and monomers at meiosis II onset4. A major goal of my research program over the next five years is to understand the molecular mechanisms underlying this remarkable process. Additionally, we will decipher the signals and pathways that trigger the formation of Rim4 assemblies and we will elucidate how translation is regulated by these structures. Budding yeast is a powerful experimental system to study these processes. My lab can easily grow populations of cells that rapidly and synchronously produce, and then clear, amyloid-like Rim4. We have developed fluorescence-based reporters that allow us to quantify Rim4 assembly and function in single cells and selection-based reporters that enable robust genetic screens. We have used these tools, in combination with biochemical approaches, to begin to illuminate the essential factors that govern how these extraordinary structures are constructed by the cell and the molecular mechanisms underpinning their function. The MIRA award provided me with the support that I needed to launch and maintain my independent research program. While we made substantial progress towards our stated goals, this award also gave the researchers in my lab the freedom to explore fruitful departures from our main focus. For example, we discovered that yeast cells utilize amyloid-like Rim4 to block the translation of retrotransposon mRNA to protect the genomic integrity of developing gametes. By making these unexpected discoveries and by developing new tools, my lab has entered an exciting phase where we are rapidly gaining important insights into how cells regulate, utilize, and process amyloid-like assemblies. Each discovery serves as a potential lead toward a therapy for amyloid-related diseases, which are needed because few, if any, effective preventative therapies are currently available. Columbia University and the Taub Institute for Alzheimer’s and Aging Research provides the supportive framework and collaborative opportunities to make this possible.

项目摘要 尽管淀粉样蛋白在许多人类疾病的病因学中的作用被最好地理解， 病理性蛋白质沉积、淀粉样蛋白和淀粉样蛋白组装体可发挥关键功能1。调解 这些功能，一些细胞具有调节这些结构的组装和清除的能力2，3。我 研究愿景是描绘健康细胞进化的生理途径和机制， 调节淀粉样蛋白的形成、功能和可逆性。这些研究的动机是我们发现， 为了控制减数分裂期间的翻译，芽殖酵母调节RNA结合蛋白Rim 4的组装 转化为具有淀粉样蛋白（因此称为淀粉样蛋白样）的几种生化特性的翻译阻遏物2。 从这些研究中获得的知识将导致我们对疾病原因的理解取得重要进展。 神经退行性疾病，并及时可能导致治疗的机会。 基于我们最近研究Rim 4清除的进展，我们现在认识到酵母细胞使用多- 位点磷酸化，在减数分裂II开始时将淀粉样Rim 4突然逆转为寡聚体和单体4。一 我的研究计划在未来五年的主要目标是了解分子机制 这一非凡过程的基础。此外，我们还将破译触发这些信号的信号和途径。 Rim 4组装的形成，我们将阐明这些结构如何调节翻译。 芽殖酵母是研究这些过程的一个强有力的实验系统。我的实验室可以很容易地 这些细胞群快速同步地产生，然后清除淀粉样蛋白样Rim 4。我们有 开发了基于荧光的报告基因，使我们能够量化Rim 4在单细胞中的组装和功能 以及基于选择的报告基因，其能够进行稳健的遗传筛选。我们结合使用了这些工具 用生物化学的方法，开始阐明控制这些非凡的 结构是由细胞和支持其功能的分子机制构建的。 MIRA奖为我提供了支持，我需要启动和保持我的独立 研究计划。虽然我们在实现既定目标方面取得了实质性进展，但该奖项也为我们提供了 我实验室的研究人员可以自由地探索我们主要关注的富有成效的偏离。比如我们 发现酵母细胞利用淀粉样蛋白Rim 4阻断反转录转座子mRNA的翻译， 发育中的配子的基因组完整性通过这些意想不到的发现和开发新的 工具，我的实验室已经进入了一个令人兴奋的阶段，我们正在迅速获得重要的见解，细胞如何 调节、利用和加工淀粉样蛋白样组装体。每一个发现都是一个潜在的线索， 淀粉样蛋白相关疾病的治疗，这是必要的，因为很少，如果有的话，有效的预防性治疗 目前可用。哥伦比亚大学和陶布老年痴呆症和衰老研究所 提供了支持性框架和合作机会，使之成为可能。