Structure and function of the portal vertex on the herpes simplex virus capsid

单纯疱疹病毒衣壳汇管顶点的结构和功能

• 批准号: 10592314
• 负责人: James F. Conway
• 金额: $ 65.25万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592314
• 关键词: Architecture; Binding; Biochemical; Capsid; Cell Separation; Complex; Cryoelectron Microscopy; DNA Packaging; Data; Defect; Development; Genes; Genetic; Genome; Herpesviridae; Herpesviridae Infections; Herpesvirus 1; Image; In Situ; Infection; Infection prevention; Knowledge; Laboratories; Length; Morphology; Positioning Attribute; Process; Proteins; Publishing; Reaction; Research; Role; Simplexvirus; Site; Structure; Techniques; Testing; Therapeutic; Therapeutic Effect; Viral; Viral Genes; Viral Genome; Viral Packaging; Viral Proteins; Virion; Virus; Virus Replication; Visualization; adenovirus penton protein; density; design; human pathogen; innovation; insight; mutant; prevent; protein function; reconstruction; recruit; structural biology; terminase; viral DNA; virology

ABSTRACT This project aims to elucidate the interactions of the herpes simplex virus portal vertex with replicated viral DNA and the HSV packaging machinery. The packaging and cleavage of replicated viral DNA into preformed capsids is a critical step during herpesvirus infection. For herpes simplex virus type 1 (HSV-1), this process requires the products of seven viral genes: UL6, UL15, UL17, UL25, UL28, UL32, and UL33. Most of the proteins involved in capsid assembly and DNA packaging are conserved suggesting that these mechanisms will also be similar for all herpesviruses. The proposed research by the Homa and[CJF1] Conway laboratories combines expertise in virology and structural biology for determining the interaction of the cleavage/packaging proteins with the capsid portal vertex leading to stable packaging of a unit length genome. Understanding the herpesvirus portal structure and its interaction with the packaging machinery has been hampered due to the subtle morphological differences between the portal and penton vertices. However, recent advances in cryoEM techniques have allowed portals to be localized in images of intact capsids yielding important structural information about the portal and associated proteins. The pUL6 portal dodecamer appears to be anchored to the capsid by interactions with the peri-pentonal triplexes as well as helical density attributed to the pUL17 and pUL25 subunits of the capsid vertex specific component (CVSC). Here we propose combined biochemical and structural studies focusing on the unique portal vertex of wild type HSV and packaging mutants that will provide insights into the mechanism of HSV genome packaging. We aim to understand the functions of the CVSC components, their interactions with the portal vertex and with the terminase complex in cleavage and retention of the packaged viral genome. Based on published and preliminary studies, we believe the pUL17 is emerging as a key player in capsid assembly and maturation. We hypothesize that pUL17 anchors the terminase complex to the portal and, after packaging is complete, recruits pUL25 to the portal to retain the viral genome. We will test the hypothesis in two specific Aims that together investigate the structure and function of the portal vertex. Aim 1 focuses on the viral proteins required for the portal complex to interact with the replicated viral genome and initiate the cleavage packaging reaction. Aim 2 follows a complementary structural approach to understand the architecture of the portal vertex, include the components required for different stages of assembly and packaging. Our results will inform the development of innovative and specific therapeutics designed to prevent herpesvirus replication by blocking DNA packaging.

摘要 该项目旨在阐明单纯疱疹病毒门脉顶点与 复制病毒DNA和HSV包装机械。蛋白质的包装和切割 将病毒DNA复制到预先形成的衣壳中是疱疹病毒感染过程中的关键步骤。为 单纯疱疹病毒1型(HSV-1)，这一过程需要七种病毒基因的产物： UL6、UL15、UL17、UL25、UL28、UL32和UL33。大多数参与衣壳组装的蛋白质 DNA包装是保守的，这表明这些机制对所有人来说也将是相似的 疱疹病毒。霍马实验室和[CJF1]康威实验室拟议的研究结合 病毒学和结构生物学方面的专门知识，用于确定病毒的相互作用 具有衣壳入口顶点的切割/包装蛋白质导致单位的稳定包装 长度基因组。了解疱疹病毒门户结构及其与病毒的相互作用 包装机械一直受到阻碍，因为它们之间的细微形态差异 门户和五角星顶点。然而，低温电磁技术的最新进展使 门户将在完整衣壳的图像中定位，从而产生关于 门户及相关蛋白。PUL6门户十二聚体似乎锚定在 衣壳与周五角形三联体的相互作用以及归因于 衣壳顶点特定成分(CVSC)的pUL17和pUL25亚基。在这里，我们建议 野生型独特门脉顶端的生化与结构相结合的研究 HSV和包装突变体将为深入了解HSV基因组的机制提供帮助 包装。我们的目标是了解CVSC组件的功能、它们与 门静脉顶端和末端酶复合体对包装病毒的切割和滞留 基因组。根据已发表的和初步的研究，我们认为pUL17正在成为一个关键 球员在衣帽的组装和成熟。我们假设pUL17锚定了末端酶 复杂到门户，在打包完成后，将pUL25招募到门户以保留 病毒基因组。我们将在两个特定目标中测试该假设，这两个目标一起调查 门户顶点的结构和功能。目标1集中在病毒所需的蛋白质上 与复制的病毒基因组相互作用并启动切割包装的门户复合体 反应。目标2遵循补充的结构方法来理解 门户顶点，包括组装的不同阶段所需的组件 包装。我们的结果将为创新和特定疗法的发展提供信息 旨在通过阻止DNA包装来防止疱疹病毒复制。