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Herpes Simplex Virus Capsid Assembly and DNA Packaging

单纯疱疹病毒衣壳组装和 DNA 包装

基本信息

批准号：
7737371
负责人：
FREDERICK L HOMA
金额：
$ 31.51万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-12-01 至 2011-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7737371
关键词：
Affect Affinity Chromatography Animals Antiviral Agents Baculoviruses Binding Biochemical Biochemical Genetics Biology Capsid Capsid Proteins Cell physiology Cells Complex Cytomegalovirus DNA DNA Binding DNA Packaging DNA Sequence Development Disease Genes Genetic Genome Goals Herpesviridae Herpesvirus 1 Herpesvirus Type 3 Homologous Gene Human Human Herpesvirus 4 Hybrids Immune Immunocompromised Host In Vitro Individual Infection Left Length Location Maps Medical Minor Mutation Newborn Infant Nucleic Acids Population Process Production Protein Binding Proteins Proteomics Reaction Research Research Personnel Role Series Simplexvirus Site Specificity Staging System Tertiary Protein Structure Testing Translations Viral Viral Proteins Yeasts insertion/deletion mutation mutant novel prevent programs protein complex protein degradation protein function protein protein interaction terminase viral DNA yeast two hybrid system

项目摘要

Infections with human herpesviruses are endemic in the population with some herpesviruses causing severe disease, especially in immune impaired individuals and newborns. The increased immunocompromised population has created an unmet medical need for antivirals against herpesviruses. The goal of the proposed project is to gain a better understanding of DNA cleavage and packaging in herpes simplex virus (HSV) through the analysis of two viral proteins, UL25 and UL28, that are required for this process. The specific hypothesis behind the proposed research is that UL25 and UL28 are required for packaging DNA into HSV capsids at separate stages in the cleavage packaging reaction. The UL28 protein is required for cleavage of concatemeric DNA into unit length molecules while UL25 serves an essential function in the production of DNA containing capsids. Although, the focus will be on UL25 and UL28 the goal is to characterize the cleavage/packaging process using genetic and biochemical approaches to examine how mutations in the UL25 and UL28 genes affect the interaction of these proteins with: (i) DNA, (ii) capsid proteins, (iii) cleavage/packaging proteins and (iv)host cell proteins. Specific goals of the project are to: 1. Determine the capsid location, DNA binding specificity and protein-protein interactions of UL25. 2. Identify functional domains of UL28 that are important for capsid incorporation, binding viral DNA packaging sites and for interaction with terminase (UL15) and portal protein (UL6) by characterizing a series of HSV-1 UL28 mutants that we have isolated; and map the second site mutation for a revertent of one of the lethal UL28 linker- insertion mutants since marker rescue and DNA sequencing have demonstrated that the mutation does not map to the UL28 gene. 3. Examine the interaction of UL25 and UL28 with viral and cell proteins using proteomic and biochemical approaches in order to identify essential protein-protein interactions and protein complexes that are involved in the cleavage/packaging reaction. These studies aim to elucidate the mechanism underlying HSV DNA cleavage and packaging and may suggest novel targets for the development of antivirals.

人类疱疹病毒感染在人群中流行，某些疱疹病毒会导致严重的症状疾病，尤其是免疫受损的个体和新生儿。免疫功能低下者增加人口对疱疹病毒抗病毒药物的医疗需求尚未得到满足。拟议的目标该项目旨在更好地了解单纯疱疹病毒 (HSV) 中的 DNA 切割和包装通过分析该过程所需的两种病毒蛋白 UL25 和 UL28。具体的这项研究背后的假设是 UL25 和 UL28 是将 DNA 包装到 HSV 中所必需的衣壳在裂解包装反应的不同阶段。裂解需要 UL28 蛋白将 DNA 串联成单位长度的分子，而 UL25 在生产中发挥着重要作用含有衣壳的DNA。虽然重点将放在 UL25 和 UL28 上，但目标是表征使用遗传和生化方法的切割/包装过程来检查突变如何发生 UL25 和 UL28 基因影响这些蛋白质与以下物质的相互作用：(i) DNA、(ii) 衣壳蛋白、(iii) 切割/包装蛋白和（iv）宿主细胞蛋白。该项目的具体目标是： 1. 确定 UL25 的衣壳定位、DNA 结合特异性和蛋白质-蛋白质相互作用。 2. 识别功能 UL28 的结构域对于衣壳掺入、结合病毒 DNA 包装位点和通过表征一系列 HSV-1 UL28 突变体与终止酶 (UL15) 和门户蛋白 (UL6) 的相互作用我们已经隔离了；并绘制致命 UL28 连接子之一的回复体的第二个位点突变- 插入突变体，因为标记救援和 DNA 测序已证明该突变不会映射到 UL28 基因。 3. 使用以下方法检查 UL25 和 UL28 与病毒和细胞蛋白的相互作用蛋白质组学和生化方法，以确定重要的蛋白质-蛋白质相互作用和蛋白质参与裂解/包装反应的复合物。这些研究旨在阐明 HSV DNA 切割和包装的潜在机制，并可能为 HSV DNA 切割和包装提供新的靶点抗病毒药物的开发。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structure and polymorphism of the UL6 portal protein of herpes simplex virus type 1.

1型单纯疱疹病毒UL6门户蛋白的结构和多态性。

DOI：
10.1128/jvi.78.22.12668-12671.2004
发表时间：
2004
期刊：
Journal of virology
影响因子：
5.4
作者：
Trus,BenesL;Cheng,Naiqian;Newcomb,WilliamW;Homa,FredL;Brown,JayC;Steven,AlasdairC
通讯作者：
Steven,AlasdairC

Major capsid reinforcement by a minor protein in herpesviruses and phage.

在疱疹病毒和噬菌体中，次要蛋白质的主要衣壳加固。