Icosahedral virion assembly and function

二十面体病毒粒子的组装和功能

• 批准号: 1408217
• 负责人: Bentley Fane
• 金额: $ 100万
• 依托单位: University of Arizona
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1408217&HistoricalAwards=false
• 关键词: Icosahedral; virion; assembly; function

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.

该项目的主要目标是了解病毒如何组装和运输其遗传物质（基因组）进入细胞。 当细胞被病毒感染时，细胞机制被用来产生病毒构建模块（蛋白质），这些模块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。 对于具有由单链DNA组成的基因组的微病毒，机器包含能量和指令，以形成管状DNA递送装置，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，它福尔斯。 破译这种基于生物学的管状输送装置的构建和功能的机制将在纳米技术和蛋白质工程领域得到应用。 该项目还将通过接触参加主要研究员教授的实验室课程的学生而产生教育影响。 部分拟议的研究将在病毒学实验室课程中进行，学生将进行假设驱动的研究。本课程的教学模式已经被证明是成功的，产生了两个与本科生合著的课堂论文。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phiX 174通过DNA先导蛋白H的寡聚化瞬时产生管，DNA通过该管从病毒衣壳递送到宿主细胞。 在DNA递送之后，管分解。该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从病毒运输到宿主，哪些因素有助于使管正确长度，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，并且这些研究将通过在体外重建早期病毒组装步骤来扩展。 由于其他病毒可能以与phiX 174相同的方式运作，因此这项研究的结果应该为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相互作用提供普遍的见解。

项目成果

ϕX174 Procapsid Assembly: Effects of an Inhibitory External Scaffolding Protein and Resistant Coat Proteins In Vitro

X174 衣壳组装：抑制性外部支架蛋白和抗性外壳蛋白的体外作用

• DOI: 10.1128/jvi.01878-16
• 发表时间: 2016
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Cherwa, James E.;Tyson, Joshua;Bedwell, Gregory J.;Brooke, Dewey;Edwards, Ashton G.;Dokland, Terje;Prevelige, Peter E.;Fane, Bentley A.;Sandri-Goldin, Rozanne M.
• 通讯作者: Sandri-Goldin, Rozanne M.

Coat Protein Mutations That Alter the Flux of Morphogenetic Intermediates through the ϕX174 Early Assembly Pathway

外壳蛋白突变通过 ÏX174 早期组装途径改变形态发生中间体的通量

• DOI: 10.1128/jvi.01384-17
• 发表时间: 2017
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Blackburn, Brody J.;Li, Shuaizhi;Roznowski, Aaron P.;Perez, Alexis R.;Villarreal, Rodrigo H.;Johnson, Curtis J.;Hardy, Margaret;Tuckerman, Edward C.;Burch, April D.;Fane, Bentley A.
• 通讯作者: Fane, Bentley A.

Elevating fitness after a horizontal gene exchange in bacteriophage φX174

噬菌体水平基因交换后提高适应性 ÏX174

• DOI: 10.1016/j.virol.2016.10.029
• 发表时间: 2017
• 期刊: Virology
• 影响因子: 3.7
• 作者: Doore, Sarah M.;Schweers, Nicholas J.;Fane, Bentley A.
• 通讯作者: Fane, Bentley A.