Architecture of endolysosomal pathways in tetrahymena

四膜虫内溶酶体途径的结构

• 批准号: 1937326
• 负责人: Aaron Turkewitz
• 金额: $ 87.13万
• 依托单位: University of Chicago
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1937326&HistoricalAwards=false
• 关键词: Architecture; endolysosomal; pathways; tetrahymena

The project will address fundamental questions about the evolution of the relationship between structure and function in cells. All cells share a set of activities, many of which involve interactions with their environment. The subject of this proposal is the pathways that allow cells to take up particles from their environment, which can be carried to a compartment where they are digested. Alternatively, such ingested particles can be transported to other compartments and integrated into cellular metabolism. The research will determine if different cells are relying on the same mechanisms, or whether, over time, have evolved diverse mechanisms to achieve similar tasks. The research will also address how cells control the transport of ingested particles by implementing experiments which test the hypothesis that the various possible outcomes are in competition with one another, and that a cell can precisely regulate the “flow” down different pathways. The project will be pursued in the ciliate Tetrahymena thermophila and will focus on pathways in the ‘endolysosomal’ network. Tetrahymena ingests bacteria by a phagocytosis-dependent pathway, and then digests those bacteria in phagolysosomes whose activity depends on delivery of hydrolytic enzymes. The research will define the pathways involved in delivery of proteins to phagolysosomes and ask whether they are the same or different as the pathways used in mammalian cells. In Tetrahymena, hydrolytic enzymes are also delivered to generate a specific secretory compartment. Experiments in the project will test the hypothesis that competition between molecular tethering complexes in the CORVET family determines the relative flux of key hydrolytic enzymes to secretory organelles vs phagolysosomes, depending on cellular conditions. The experiments will be based on natural and induced fluctuations in the abundance of specific CORVET subunits. The experiments will also take advantage of the fact that CORVET tethers are recruited by Rab GTPases, so that manipulation of Rab activities can be used to indirectly influence CORVET function. The project will provide an environment to introduce undergraduates to research, and for the training of one PhD student and one postdoctoral fellow.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.

该项目将解决有关细胞中结构与功能之间关系演变的基本问题。所有细胞共享一组活动，其中许多活动涉及与环境的相互作用。该提案的主题是允许细胞从其环境中占据颗粒的途径，可以将其带到被消化的隔室。或者，这种摄入的颗粒可以运输到其他隔室并整合到细胞代谢中。该研究将确定不同的细胞是否依赖相同的机制，或者随着时间的流逝是否发展了潜水机制以实现相似的任务。这项研究还将通过实施实验来解决细胞如何控制摄入的颗粒的运输，从而测试各种可能的结果彼此竞争的假设，并且细胞可以准确地调节不同的途径的“流动”。该项目将在纤毛四膜热嗜热菌中进行，并将集中在“内溶性”网络中的途径上。四膜虫通过吞噬作用依赖性途径摄入细菌，然后消化这些细菌在吞噬体中的活性取决于水解酶的递送。该研究将定义蛋白质递送至吞噬物体的途径，并询问它们与哺乳动物细胞中使用的途径是否相同或不同。在四膜虫中，还递送水解酶以产生一个特定的秘密室。该项目中的实验将检验以下假设：Corvet家族中分子束缚复合物之间的竞争决定了关键水解酶的相对通量，取决于细胞状况，取决于细胞条件。该实验将基于特定Corvet亚基的抽象中的自然和诱导的波动。实验还将利用Rab GTPases募集Corvet Teathers的事实，因此可以使用RAB活动的操纵来间接影响Corvet功能。该项目将提供一个环境，以介绍本科生进行研究，并为一名博士学位学生和一名博士后研究员进行培训。该奖项反映了NSF的法定任务，并通过使用该基金会的知识分子优点和更广泛的影响评估标准来评估NSF的法定任务。