Identification and functional characterization of novel molecular components involved in cell motility: Taking advantage of bacterial pathogens

参与细胞运动的新型分子成分的鉴定和功能表征：利用细菌病原体

• 批准号: RGPIN-2018-05100
• 负责人: Guttman, Julian
• 金额: $ 3.64万
• 依托单位: Simon Fraser University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=713501
• 关键词: Identification; functional; characterization; novel; molecular

Summary of proposal to the public Cellular movement is crucial for organism development, wound healing, and tissue maintenance. When cells migrate, they use their internal skeleton, called the cytoskeleton, in dynamic ways. The cytoskeleton is made-up of a number of components, but by far the most important for cell motility is the actin cytoskeleton. Actin proteins form filaments that assemble and disassemble. When coordinated with the periphery of the cell, at the cell membrane, these force-generating actions enable the cell to move. Because of the complexity of the events occurring at the front of a migrating cell, researchers have identified other actin-based motility systems that mimic the actions occurring during whole cell movement. Some of the most often used model systems exploit microbes that enter cells and hijack the actin cytoskeleton of those cells during their infectious processes. The control of the actin cytoskeleton causes the bacteria or viruses to use the actin within their hosts for their own movement. One of these microbes, pathogenic E. coli, dock onto the outside of the cells that they infect and generate small structures referred to as “actin pedestals” at regions of E. coli attachment. These pedestals enable the bacteria to “surf” atop the infected cells and provide an ideal system to identify new proteins used during actin-based cellular motility. Using the funds from our previous NSERC grant we developed a strategy to concentrate E. coli pedestals and used mass spectrometry to identify >90 proteins that had never been identified at these sites, we have begun the process of characterizing these proteins to determine their functions at various actin-rich structures and during whole cell motility. We expect that our work will provide new cellular proteins involved in actin/cell membrane interactions as well as proteins that associate with actin itself and regulate its functions.

向公众提议的摘要 细胞运动对于生物体发育，伤口愈合和维持组织至关重要。当细胞迁移时，他们会以动态方式使用其内部骨架，称为细胞骨架。细胞骨架由许多成分组成，但到目前为止，对于细胞运动，最重要的是肌动蛋白细胞骨架。肌动蛋白蛋白会形成组装和拆卸的细丝。当与细胞的外围物协调时，在细胞膜上，这些力产生的作用使细胞能够移动。由于事件发生在迁移细胞正面的事件的复杂性，研究人员已经确定了其他基于肌动蛋白的运动系统，这些系统模仿了整个细胞运动过程中发生的动作。一些经常使用的模型系统利用了进入细胞的微生物，并在传染过程中劫持了这些细胞的肌动蛋白细胞骨架。肌动蛋白细胞骨架的控制会导致细菌或病毒在其宿主中使用肌动蛋白进行自身运动。这些微生物之一是致病性大肠杆菌，将它们靠近它们感染的细胞外部，并在大肠杆菌附着区域产生称为“肌动蛋白基座”的小结构。这些基座使细菌能够在感染细胞上“浏览”，并提供理想的系统，以鉴定基于肌动蛋白的细胞运动过程中使用的新蛋白质。使用以前的NSERC赠款中的资金，我们制定了一种集中大肠杆菌基座的策略，并使用质谱法来识别从未在这些地点鉴定的> 90个蛋白质，我们已经开始表征这些蛋白质以确定其在各种肌动蛋白富含结构和整个细胞运动中的功能。我们预计我们的工作将为肌动蛋白/细胞膜相互作用以及与肌动蛋白本身相关并调节其功能的蛋白质提供新的细胞蛋白。