Assembly of the Herpes Simplex Virus Capsid

单纯疱疹病毒衣壳的组装

• 批准号: 9417770
• 负责人: Jay Brown
• 金额: $ 31.84万
• 依托单位: University of Virginia Main Campus
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9417770&HistoricalAwards=false
• 关键词: Assembly; Herpes; Simplex; Virus; Capsid

The capsid of herpes simplex virus type 1 (HSV 1) is an icosahedral (T=16) protein shell 15nm thick and 125nm in diameter. In the intact virus, the capsid contains the viral DNA and it is surrounded by a lipid membrane. The capsid is composed of a major protein species called VP5 which accounts for approximately 70% of the total capsid mass plus three minor polypeptides, VP19, VP23 and VP26. In infected cells, empty (DNA free) capsids are assembled in the cell nucleus and later packaged with DNA. The P.I. has recently developed a system in which the HSV 1 capsid can be assembled in a cell free environment. The system requires the use of six different recombinant baculoviruses each coding for one of the six proteins involved in HSV 1 capsid formation. The six are the four structural polypeptides mentioned above plus the scaffolding protein (VP22a) and the maturational protease (the product of the HSV 1 UL26 gene). Each virus is used to infect a separate culture of insect (Sf9) cells and cell free extracts are prepared from each culture. HSV 1 capsids form spontaneously when the six extra cts are mixed. The P.I. plans to study the time course of capsid formation in this system to identify morphogenetic intermediates in the assembly process. It is expected that the structure and composition of such intermediates and the times they appear in reaction mixtures will suggest the basic features of the overall assembly pathway. %%% Herpes simplex virus type 1 (HSV l) is the causative agent of cold sores and fever blisters. The infectious virus consists of a protein shell called the capsid surrounded by a lipid membrane. HSV l contains DNA which is found inside the capsid shell. When the virus is replicated in an infected cell, a complete, empty capsid is first formed; it is later packaged with DNA and finally coated with a membrane envelope. The goal of this project is to understand the first step in HSV l assembly, namely formation of the capsid. In these studies, the six proteins involved in capsid assembly are mixed together in vitro under conditions in which capsids form spontaneously. The P.I. proposes to use this system to identify intermediates in the assembly process. It is expected that the structure and protein composition of such intermediates and the times they appear will suggest the basic features of the overall assembly pathway.

单纯疱疹病毒1型（HSV 1）的衣壳是一个二十面体（T=16）蛋白壳，厚15 nm，直径125 nm。在完整的病毒中，衣壳含有病毒DNA，并被脂质膜包围。衣壳由称为VP 5的主要蛋白质种类（占总衣壳质量的约70%）加上三种次要多肽（VP 19、VP 23和VP 26）组成。在受感染的细胞中，空（无DNA）衣壳在细胞核中组装，随后与DNA包装在一起。私家侦探最近开发了一种系统，其中HSV 1衣壳可以在无细胞环境中组装。该系统需要使用六种不同的重组杆状病毒，每种重组杆状病毒编码参与HSV 1衣壳形成的六种蛋白中的一种。这六种是上述四种结构多肽加上支架蛋白（VP 22 a）和成熟蛋白酶（HSV 1UL 26基因的产物）。每种病毒用于感染昆虫（Sf 9）细胞的单独培养物，并从每种培养物制备无细胞提取物。当六个额外的cts混合时，HSV 1衣壳自发形成。私家侦探计划研究在这个系统中衣壳形成的时间过程，以确定组装过程中的形态发生中间体。预计这些中间体的结构和组成以及它们在反应混合物中出现的时间将表明整个组装途径的基本特征。 %%% 单纯疱疹病毒1型（HSV 1）是唇疱疹和发热性水泡的病原体。感染性病毒由一个被称为衣壳的蛋白质外壳和一层脂质膜组成。HSV 1含有在衣壳壳内发现的DNA。当病毒在受感染的细胞中复制时，首先形成一个完整的空衣壳;然后用DNA包装，最后包被一层膜被。该项目的目标是了解HSV 1组装的第一步，即衣壳的形成。在这些研究中，在衣壳自发形成的条件下，将参与衣壳组装的六种蛋白质在体外混合在一起。私家侦探建议使用该系统来识别组装过程中的中间体。预计这些中间体的结构和蛋白质组成以及它们出现的时间将表明整个组装途径的基本特征。