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，细胞机制被用来制造病毒构件（蛋白质），这些蛋白质与基因组组装成成熟的病毒，这些病毒可以被认为是准备感染其他细胞的小机器。对于基因组由单链DNA组成的微病毒来说，这些机器包含能量和指令，以形成管状DNA传递装置，将DNA从病毒运输到宿主细胞。只有当病毒在被感染的细胞表面时，管才会形成，一旦DNA被传递，管就会破裂。破译这种基于生物的管状输送装置的结构和功能的机制将在纳米技术和蛋白质工程领域具有应用价值。该项目还将通过招收由首席研究员教授的实验课程的学生，对教育产生影响。部分建议的研究将在病毒学实验课程中进行，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，产生了两篇由本科生共同撰写的班级出版物。传染性病毒的产生需要一系列复杂的大分子相互作用。无尾的二十面体细菌病毒phiX174通过DNA先导蛋白H的寡聚化瞬间产生一个管状结构，DNA通过这个管状结构从病毒衣壳传递到宿主细胞。DNA传递后，试管解体。这一建议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，以了解该导蛋白的化学组成如何使其能够将DNA从病毒运输到宿主，哪些因素有助于使管的长度正确，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构早期病毒组装步骤来扩展。由于其他病毒可能以与phiX174相同的方式运作，本研究的结果应提供对无尾病毒组装和感染重要的蛋白质-蛋白质相互作用和蛋白质- dna相互作用的一般见解。

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.