Development of Yeast Prionomics

智力价值：自组装的纤维蛋白聚集体(淀粉样蛋白)可能代表着一个古老的蛋白质折叠。淀粉样蛋白的种子聚合为淀粉样蛋白状态的潜在传染性或遗传性提供了基础。事实上，最近的证据表明，形成可传递的淀粉样蛋白(Prion)的能力在酵母蛋白中普遍存在，很可能在其他生物体的蛋白质中也是如此。在遗传和进化过程中，我们不再可能忽视Pron的潜在输入。然而，对酵母蛋白的生物学作用的了解仍处于初级水平，部分原因是缺乏独立于序列和表型的检测和监测蛋白的方法。这项研究的总体目标是组成酵母普里恩组，也就是能够在其天然状态下形成普鲁恩的蛋白质目录，并评估对酵母生物学和进化的影响。这将有助于确定酵母菌株的Pron图谱是否像基因模式一样是由自然选择驱动的。为此，人们正在开发独立于菌株基因并适用于高通量分析和Pron图谱的Pron检测方法。具体的研究目标如下：1)优化非序列生物化学方法快速检测普恩病毒；2)鉴定新的普恩候选基因。智力优势的驱动因素是对基于蛋白质的遗传这一迅速兴起的主题的重视(这一主题仍然被严重地研究不足，可能会显著改变我们对生物进化的理解)。新的无偏生化工具正在被开发并应用于重要的生物学问题，如鉴定新的Prion，表征它们的作用，以及确定酵母菌株的完整的Prion图谱。更广泛的影响：用于检测淀粉样蛋白和普恩蛋白的生化方法和工具有可能适用于高通量分析，并可用于表征酵母和其他生物在其自然环境中的主序。这将打开一个全新的环境“原始经济学”领域，为环境基因组学提供补充，并可能对理解淀粉样蛋白的生物学作用以及被假设涉及类普里子开关的过程产生深远的影响，如记忆、压力保护和细胞内结构的组装。在该项目过程中开发的新的淀粉样蛋白检测工具，以及关于糖利用和普里恩之间的联系的数据，可能会对生物技术行业感兴趣。该项目在很大程度上依赖学生的参与，并将与教育进程紧密结合，为博士论文以及本科生和研究生课程提供学习科目。PI的实验室参与了专注于大分子组装和分子进化的跨学科中心，这些中心将从该项目的成功中受益匪浅。

Intellectual Merit: Self-assembled fibrous protein aggregates (amyloids) probably represent an ancient protein fold. Seeded polymerization of an amyloid provides a basis for potential infectivity or heritability of an amyloid state. Indeed, recent evidence demonstrates that ability to form transmissible amyloids (prions) is widespread among yeast proteins, and likely among the proteins from other organisms. It is no longer possible to ignore the potential input of prions in inheritance and evolution. However, understanding of the biological roles of yeast prions remains at rudimentary levels, in part due to the lack of sequence- and phenotype-independent approaches for prion detection and monitoring. Overall goal of this research is to compose the yeast prionome, that is, a catalogue of proteins capable of forming prions in their native state, and to assess impact on yeast biology and evolution. This will help to determine if prion profiles of the yeast strains are driven by natural selection in the same way as genotypic patterns are. For this purpose, approaches for prion detection are being developed that are independent of strain genotype and applicable to the high throughput analysis and prion profiling. Specific research objectives are as follows: 1) to optimize the sequence-independent biochemical approaches for rapid prion detection; 2) to characterize new prion candidates. Intellectual merit is driven by the emphasis on a rapidly emerging topic of protein-based inheritance (that is still grossly understudied and may significantly change our understanding of biological evolution). New unbiased biochemical tools for prion detection are being developed and applied to important biological questions, such as identification of new prions, characterization of their effects, and determining of the complete prion profiles of the yeast strains. Broader Impacts: Biochemical approaches and tools for detection of amyloids and prions are potentially amenable to high throughput analysis and can be applied to characterizing prionomes of yeast and other organisms in their natural environments. This will open a whole new area of environmental "prionomics", providing a complement to environmental genomics, and may have far-reaching implications for understanding the biological roles of amyloids and the processes that have been hypothesized to involve prion-like switches, such as memory, protection from stresses, and assembly of intracellular structures. New amyloid detection tools, developed in the course of this project, and data on the connection between the sugar utilization and prions could be of interest to the biotechnological industry. This project heavily relies on participation of students and will be tightly integrated with the educational process by providing subjects of study for Ph.D. dissertations and for teaching undergraduate and graduate courses. The PI's lab participates in the interdisciplinary centers focused on macromolecular assemblies and molecular evolution, and these centers will significantly benefit from the success of the project.