Icosahedral virion assembly and function

该项目的主要目标是了解病毒如何组装并将其遗传物质(基因组)运输到细胞中。当细胞被病毒感染时，细胞机器被用来产生病毒构建块(蛋白质)，这些病毒与基因组组装成成熟的病毒，可以被认为是准备感染其他细胞的小机器。微型病毒的基因组由单链DNA组成，机器含有能量和指令，可以形成管状DNA输送装置，将DNA从病毒输送到宿主细胞。只有当病毒在被感染的细胞表面时，管子才会形成，一旦DNA被传递，它就会解体。破译这种基于生物的管状输送装置的结构和功能的潜在机制将在纳米技术和蛋白质工程领域得到应用。该项目还将对参加首席调查员讲授的实验室课程的学生产生教育影响。部分拟议的研究将在病毒学实验室课程中进行，在该课程中，学生将进行假设驱动的研究。这门课程的教育模式已经被证明是成功的，产生了两本与本科生共同撰写的课堂出版物。传染性病毒的产生需要一系列复杂的大分子相互作用。无尾二十面体细菌病毒phiX174通过DNA引导蛋白H的寡聚瞬间产生一管，DNA通过该管从病毒衣壳输送到宿主细胞。DNA输送后，试管就会解体。这项提案旨在解决有关病毒组装和感染的几个重要问题。遗传和结构方法的结合将探索H蛋白的结构-功能关系，以了解这种先导蛋白的化学组成如何使其能够将DNA从病毒运送到宿主，哪些因素有助于使管子具有合适的长度，以及哪些其他蛋白质与H相互作用以促进适当的功能。其他实验将探索H蛋白如何与病毒衣壳内的内部支架蛋白相互作用，以促进成熟病毒的组装，这些研究将通过在体外重组早期病毒组装步骤来扩展。由于其他病毒可能以与phiX174相同的方式工作，因此这项研究的结果应该会为蛋白质-蛋白质相互作用和蛋白质-DNA相互作用提供一般性的见解，这些相互作用对于组装和感染无尾病毒很重要。

The main goal of this project is to understand how viruses assemble and transport their genetic material (genome) into cells. When cells are infected by viruses, the cellular machinery is used to produce viral building blocks (proteins) that assemble with the genome into mature viruses that can be thought of as little machines ready to infect other cells. For microviruses, which have genomes made up of single-stranded DNA, the machines contain energy and instructions to form a tubular DNA delivery device to transport DNA from the virus to a host cells. The tube only forms when the virus is on the surface of the cell to be infected, and it falls apart once the DNA has been delivered. Deciphering the mechanisms underlying construction and function of this kind of biologically-based tubular delivery device will have applications to the fields of nanotechnology and protein engineering. The project will also have an educational impact by reaching students enrolled in a laboratory course taught by the principal investigator. Part of the proposed research will be conducted in a virology laboratory course in which students will conduct hypothesis-driven research. The educational paradigm for this course has already been proven successful, yielding two class-generated publications with undergraduates as co-authors.The production of infectious viruses requires a complex array of macromolecular interactions. The tail-less, icosahedral bacterial virus, phiX174, transiently produces a tube by oligomerization of a DNA pilot protein H, through which DNA is delivered from the viral capsid to the host cell. After DNA delivery, the tube disassembles. This proposal seeks to address several important questions about viral assembly and infection. A combination of genetic and structural approaches will explore the structure-function relationships in the H protein to understand how the chemical make-up of this pilot protein enables it to transport DNA from virus to host, what factors contribute to making the tube the right length, and what other proteins interact with H to promote proper function. Other experiments will explore how the H protein interacts with internal scaffolding proteins within the viral capsid to promote assembly of the mature virus, and these studies will be extended by reconstituting the early viral assembly steps in vitro. Because other viruses are likely to operate the same way as phiX174, the results of this research should provide generalized insights on the protein-protein interactions and protein-DNA interactions important for assembly and infection by tail-less viruses.