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.

这项研究将提高我们对自噬的基本理解，一种细胞清理和回收系统，有助于保护许多组织（包括人类）的细胞免受饥饿，病原体和细胞碎片的积累等压力。这项研究的结果可能导致作物产量以及动物和人类健康的提高。除了其科学目标外，该项目还将通过在东密歇根大学提供研究丰富的学习环境来帮助创建潜水员和技术识字的劳动力，这是一家小学的本科机构，为学生大致和社会经济多样化的学生提供服务。将支持十二名本科生和两名硕士学生进行1 - 2年的尖端生化研究。此外，该项目将帮助支持约24名本科生的学期指导研究经验，参加基于研究的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，技术和工程专业人士所需的实用和智力技能。该项目使用酵母遗传学来促进我们对自噬体形成的理解，自噬体是一种双重膜囊泡，它包裹了细胞货物，并将其传递给真空莫尔（或在后代，溶酶体）中以降解。尽管先前在酿酒酵母的研究已经确定了这一过程所需的蛋白质，但有关它们如何运作的运作仍有许多待了解。一个核心参与者是ATG8共轭系统，该系统将小型泛素样蛋白ATG8连接到脂质磷脂酰乙醇胺上，从而定义了形成的自噬膜。先前的研究表明，单元格中的ATG8量决定了自噬体的大大，但不是产生的数量。但是，ATG7催化导致ATG8共轭的途径中的第一步，确定了自体的大小和数量。该项目将在此途径中研究其他蛋白质，以解决此难题，并确定该途径的哪个分支影响自噬体数量。该项目还将调查ATG11的分子细节，ATG11是一种蛋白质，该蛋白通过结合并排列核心自噬蛋白来组织自噬货物周围的自噬体形成。对初步数据的ATG11区域的突变分析已证明对与其主要伙伴之一结合至关重要，将证明哪些特定残差对于这种相互作用至关重要。最后，将开发改进的数学模拟，以允许更准确的估计自动磁体大小和传输电子显微镜部分的数量。该奖项反映了NSF的法定任务，并且认为值得通过基金会的知识分子优点和更广泛的影响审查标准通过评估来获得支持。