Development of Yeast Prionomics

智力优势：自组装的纤维状蛋白质聚集体（淀粉样蛋白）可能代表了一种古老的蛋白质折叠。淀粉样蛋白的种子聚合为淀粉样蛋白状态的潜在感染性或遗传性提供了基础。事实上，最近的证据表明，形成传染性淀粉样蛋白（朊病毒）的能力在酵母蛋白中广泛存在，并且可能在其他生物体的蛋白质中存在。我们再也不能忽视朊病毒在遗传和进化中的潜在作用。然而，对酵母朊病毒的生物学作用的理解仍然处于初级水平，部分原因是缺乏序列和表型独立的朊病毒检测和监测方法。这项研究的总体目标是组成酵母朊病毒组，即能够在天然状态下形成朊病毒的蛋白质目录，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朊病毒谱是否与基因型模式一样由自然选择驱动。为此，正在开发不依赖于菌株基因型并适用于高通量分析和朊病毒谱分析的朊病毒检测方法。具体的研究目标如下：1）优化用于快速检测朊病毒的不依赖于序列的生物化学方法; 2）表征新的朊病毒候选物。知识价值是由对基于蛋白质的遗传的快速新兴主题的强调驱动的（这仍然是严重不足的研究，可能会显着改变我们对生物进化的理解）。用于朊病毒检测的新的无偏见的生物化学工具正在被开发并应用于重要的生物学问题，例如新朊病毒的鉴定，其作用的表征，以及酵母菌株的完整朊病毒谱的确定。更广泛的影响：用于检测淀粉样蛋白和朊病毒的生物化学方法和工具可能适合于高通量分析，并且可以应用于表征酵母和其他生物体在其自然环境中的朊病毒。这将打开一个全新的领域的环境“prionomics”，提供了一个补充环境基因组学，并可能有深远的影响，了解淀粉样蛋白的生物学作用和过程，已假设涉及朊病毒样开关，如记忆，保护压力，和组装的细胞内结构。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与朊病毒之间联系的数据可能会引起生物技术行业的兴趣。该项目在很大程度上依赖于学生的参与，并将通过提供博士研究课题与教育过程紧密结合。学位论文和教学本科和研究生课程。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.