Icosahedral Virion Assembly and Function

二十面体病毒粒子的组装和功能

• 批准号: 2013653
• 负责人: Bentley Fane
• 金额: $ 101.51万
• 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: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2013653&HistoricalAwards=false
• 关键词: Icosahedral; Virion; Assembly; Function

Viruses have protein shells, or capsids, that protect their genomes. During an infection, genomes must be organized, or packaged, into capsids. Genome organization is a hurdle common to all organisms. For example, the human genome is approximately 5 feet long, yet it is organized to fit within a comparatively tiny cell. A similar situation exists with viruses. The project investigates genome organization in single-stranded DNA microviruses. In this virus family, four components ensure proper packaging: the capsid, a small DNA binding protein, a gate keeper protein, and specific genome sequences. The research endeavors to identify how these components operate. In the future, this knowledge could be translated into beneficial applications, such as therapeutic viral vectors. The research contributes to the education of undergraduate, graduate and high-school students. High-school students and teachers conduct 4 to 9-month, data-generating projects, which elicit enthusiasm for science research at an earlier age. Some of the participating schools serve large populations of under-represented groups in science, which further benefits society. Single-stranded viral DNA packaging combines mechanisms found in single-stranded RNA and double-stranded DNA systems. Like double-stranded DNA, single-stranded DNA is packaged into preformed shells but like RNA genomes, they interact extensively with capsids. øX174 packaging is a four-component system: 1) the capsid’s DNA binding pockets, 2) the J protein that guides DNA between pockets, 3) specific DNA sequences, and 4) the A*(star) protein. Structural data suggest that specific coat and J protein residues order the genome into icosahedral symmetry: a hypothesis to be genetically and biochemically tested. To further elucidate protein A* function, the effects of altered protein levels and mutated A* target sites will be determined. Moreover, 60 degenerate, genomic consensus sequences may facilitate genome organization. The consensus sequences will be “de-optimized” and the resulting effects will be characterized. If “de-optimization” affects packaging, it may indicate an additional selective pressure operating a single-stranded genomes. The research outcomes will advance our understanding of the basic principles by which single-stranded DNA viruses assemble into infectious particles.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

病毒有蛋白质外壳或衣壳，可以保护它们的基因组。在感染过程中，基因组必须被组织或包装成衣壳。基因组组织是所有生物体共同的障碍。例如，人类基因组大约有5英尺长，但它被组织成适合一个相对较小的细胞。病毒也存在类似的情况。该项目研究单链DNA微病毒的基因组组织。在这个病毒家族中，有四种成分确保了正确的包装：衣壳、一种小的DNA结合蛋白、一种看门蛋白和特定的基因组序列。研究试图确定这些组件是如何工作的。在未来，这些知识可以转化为有益的应用，如治疗性病毒载体。本研究有助于本科生、研究生和高中生的教育。高中学生和教师进行4到9个月的数据生成项目，在较早的年龄激发科学研究的热情。一些参与的学校为大量科学代表性不足的群体提供服务，这进一步造福社会。单链病毒DNA包装结合了单链RNA和双链DNA系统中发现的机制。像双链DNA一样，单链DNA被包装到预先形成的外壳中，但像RNA基因组一样，它们与衣壳广泛相互作用。øX174包装是一个四组分系统：1）衣壳的DNA结合口袋，2）在口袋之间引导DNA的J蛋白，3）特定的DNA序列，和4）A*（星星）蛋白。 结构数据表明，特定的外壳和J蛋白残基顺序的基因组成二十面体对称：一个假设是遗传和生化测试。为了进一步阐明蛋白A* 功能，将确定改变的蛋白水平和突变的A* 靶位点的影响。此外，60个简并的基因组共有序列可以促进基因组组织。 共有序列将被“去优化”，并且将表征所产生的效应。如果“去优化”影响包装，则可能表明操作单链基因组的额外选择压力。该研究成果将促进我们对单链DNA病毒组装成传染性颗粒的基本原理的理解。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Low-Temperature Adaptation Targets Genome Packing Reactions in an Icosahedral Single-Stranded DNA Virus

低温适应以二十面体单链 DNA 病毒中的基因组包装反应为目标

• DOI: 10.1128/jvi.01970-21
• 发表时间: 2022
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Ogunbunmi, Elizabeth T.;Love, Samuel D.;Rhodes, Katherine A.;Morales, Adriana;Wilch, Margaret H.;Jonas, Jeremy;Fane, Bentley A.
• 通讯作者: Fane, Bentley A.

Cryo-EM Structure of Gokushovirus ΦEC6098 Reveals a Novel Capsid Architecture for a Single-Scaffolding Protein, Microvirus Assembly System

Gokushovirus δ EC6098 的冷冻电镜结构揭示了单支架蛋白、微病毒组装系统的新型衣壳结构

• DOI: 10.1128/jvi.00990-22
• 发表时间: 2022
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Lee, Hyunwook;Baxter, Alexis J.;Bator, Carol M.;Fane, Bentley A.;Hafenstein, Susan L.
• 通讯作者: Hafenstein, Susan L.

The Effects of Packaged, but Misguided, Single-Stranded DNA Genomes Are Transmitted to the Outer Surface of the ϕX174 Capsid

包装但被误导的单链 DNA 基因组的影响被传输到 X174 衣壳的外表面

• DOI: 10.1128/jvi.00883-21
• 发表时间: 2021
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Ogunbunmi, Elizabeth T.;Roznowski, Aaron P.;Fane, Bentley A.
• 通讯作者: Fane, Bentley A.