RUI: Tools and Approaches for Investigating the Basic Mechanisms of Autophagy

RUI：研究自噬基本机制的工具和方法

• 批准号: 2243163
• 负责人: Steven Backues
• 金额: $ 41.44万
• 依托单位: Eastern Michigan University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2026-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2243163&HistoricalAwards=false
• 关键词: RUI; Tools; Approaches; Investigating; Basic

This research will advance our basic understanding of autophagy, a cellular clean-up and recycling system that helps to protect cells of many organisms – including humans – from stresses such as starvation, pathogens, and the accumulation of cellular debris. Results of this research could lead to improvements in crop yields as well as animal and human health. In addition to its scientific goals, this project will help create a diverse and technically literate workforce by providing a research-rich learning environment at Eastern Michigan University, a primarily undergraduate institution that serves a racially and socioeconomically diverse population of students. Twelve undergraduate and two master’s students will be supported to perform cutting-edge biochemical research for 1-2 years each. In addition, the project will help support semester-long, guided research experiences for ~24 undergraduate students taking a research-based senior-level biochemistry laboratory course. These two activities will provide students with the practical and intellectual skills needed to become the next generation of science, technology, and engineering professionals. This project uses yeast genetics to advance our understanding of the formation of the autophagosome, a double-membraned vesicle that envelops cellular cargo and delivers it to the vacuole (or, in metazoans, the lysosome) for degradation. Although previous research in Saccharomyces cerevisiae has established the proteins necessary for this process, much remains to be learned about how they function to carry it out. One central player is the Atg8 conjugation system, which attaches the small ubiquitin-like protein Atg8 to the lipid phosphatidylethanolamine, thus defining the forming autophagic membrane. Previous research has shown that the amount of Atg8 in a cell determines how large autophagosomes become, but not the number created. However, Atg7, which catalyzes the first step in the pathway leading to Atg8 conjugation, determines both the size and the number of autophagosomes. This project will investigate additional proteins in this pathway to resolve this conundrum and determine which branch of the pathway affects autophagosome number. The project will also investigate the molecular details of Atg11, a protein that organizes autophagosome formation around autophagic cargo by binding to and arranging core autophagy proteins. Mutational analysis of a region of Atg11 that preliminary data has shown to be crucial for binding to one of its key partners will demonstrate which specific residues are essential for this interaction. Finally, an improved mathematical simulation to allow more accurate estimation autophagosome size and number from transmission electron microscopy sections will be developed.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这项研究将增进我们对自噬的基本了解，自噬是一种细胞清理和回收系统，有助于保护包括人类在内的许多生物体的细胞免受饥饿、病原体和细胞碎片积累等压力的影响。除了其科学目标外，该项目还将通过在东密歇根大学（一所主要本科院校）提供研究丰富的学习环境来帮助培养一支多元化且具有技术素养的劳动力队伍。服务于种族和社会多元化的人群将支持 12 名本科生和两名硕士生每人进行 1-2 年的尖端生化研究。此外，该项目还将帮助支持约 24 名本科生进行为期一学期的研究型研究。高级生物化学实验室课程。这两项活动将为学生提供成为下一代科学、技术和工程专业人员所需的实践和智力技能。该项目利用酵母遗传学来加深我们对自噬体形成的理解。一个包裹细胞货物并将其输送至液泡（或后生动物中的溶酶体）进行降解的双膜囊泡。 Atg8 缀合系统是其中的一个核心参与者，它将小型泛素样蛋白 Atg8 附着在脂质上。磷脂酰乙醇胺，从而定义了自噬膜的形成，之前的研究表明，细胞中 Atg8 的数量决定了自噬体的大小，而不是产生的数量。然而，Atg7 催化 Atg8 结合途径的第一步。确定自噬体的大小和数量该项目将研究该途径中的其他蛋白质以解决这一难题并确定该途径的哪个分支会影响。该项目还将研究 Atg11 的分子细节，Atg11 是一种通过结合和排列核心自噬蛋白来组织自噬体形成的蛋白质，初步数据表明该区域对于与自噬体结合至关重要。其主要合作伙伴之一将证明哪些特定残基对于这种相互作用至关重要。最后，将通过改进的数学模拟来更准确地估计自噬体的大小和数量。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。