The Cellular Actin Network and Virus Replication

这个项目将分析植物病毒如何与肌动蛋白细胞骨架相互作用，肌动蛋白细胞骨架是细胞内发生许多分子运动(运输)的蛋白质支架之一。病毒似乎以肌动蛋白网络为目标对细胞通路进行重新编程，以便它们支持病毒的生命周期，而不是执行正常的细胞功能。已经观察到，病毒会改变肌动蛋白网络的功能，从而极大地重组受感染的细胞以支持病毒复制。这项研究的潜在社会益处是更深入地了解病毒与宿主的相互作用，这可能导致有效的新的抗病毒方法。此外，这项工作将为研究生和博士后提供培训，并将为高中生提供参与指导研究的机会，并在高中科学竞赛中展示他们的结果。该项目将利用植物病毒针对细胞粘连蛋白的新发现，细胞粘连蛋白是肌动蛋白网络的主要调节因子。这项研究将解决以下问题：病毒如何利用粘连蛋白干扰肌动蛋白网络？肌动蛋白组织如何控制细胞内的病毒复制？该项目将利用酵母作为病毒的模型宿主，允许在病毒复制中测试具有良好特征的粘连蛋白和肌动蛋白突变体。对酵母和积累病毒的植物细胞中的肌动蛋白网络进行纯化成分的体外分析和基于显微镜的分析将被用来揭开病毒复制过程中受肌动蛋白影响的步骤。对病毒复制复合体的蛋白质组学分析将确定哪些被病毒破坏的宿主蛋白受肌动蛋白网络的调控。

This project will dissect how plant viruses interact with the actin cytoskeleton, one of the intracellular protein scaffolds along which much molecular movement (trafficking) occurs. Viruses appear to target the actin network to reprogram cellular pathways so that they support the virus life cycle instead of performing normal cellular functions. It has been observed that viruses modify the functionality of the actin network, thereby dramatically reorganizing the infected cells to support viral replication. A potential societal benefit of this research is a deeper understanding of virus-host interactions that could lead to potent new anti-viral approaches. In addition, this work will provide training for graduate students and a postdoctoral fellow and will provide opportunities for high school students to participate in supervised research and present their results at high school science competitions.The project will exploit the new discovery that plant viruses target cellular cofilin, a major regulator of the actin network. The research will address the questions: How does a virus exploit cofilin to interfere with the actin network? How does actin organization control viral replication in cells? The project will take advantage of yeast as a model host for viruses, allowing the testing of well-characterized cofilin and actin mutants in virus replication. In vitro assays with purified components and microscopy-based analysis of the actin network in yeast and plant cells accumulating viruses will be used to unravel the steps in the viral replication process that are affected by actin. Proteomic analysis of the viral replication complex will determine which of the host proteins that are subverted by viruses are regulated by the actin network.