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 本项目旨在阐明单纯疱疹病毒门脉顶点与 复制的病毒DNA和HSV包装机器。的包装和裂解 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），这一过程需要七个病毒基因的产物： UL 6、UL 15、UL 17、UL 25、UL 28、UL 32和UL 33。大多数参与衣壳装配的蛋白质 和DNA包装是保守的，这表明这些机制也将是相似的， 疱疹病毒Homa和[CJF 1] Conway实验室提出的研究结合了 病毒学和结构生物学的专业知识，用于确定 切割/包装蛋白质与衣壳门顶点，导致稳定包装单位 长度基因组。了解疱疹病毒的门户结构及其与 包装机械受到阻碍，由于微妙的形态差异， 入口和五邻体顶点。然而，冷冻EM技术的最新进展已经允许 门户网站被定位在图像的完整衣壳产生重要的结构信息， 门户和相关蛋白。pUL 6门脉十二聚体似乎锚定在 衣壳的相互作用，以及螺旋密度归因于 衣壳顶点特异性组分（CVSC）的pUL 17和pUL 25亚基。在这里我们建议 结合生物化学和结构研究，重点是野生型独特的门静脉顶点 HSV和包装突变体将为HSV基因组机制提供见解 包装.我们的目标是了解CVSC组件的功能，它们与 门顶点和与终止酶复合物在切割和保留包装的病毒 基因组基于已发表的和初步的研究，我们认为pUL 17正在成为一个关键的 参与衣壳的组装和成熟。我们假设pUL 17锚定末端酶， 在包装完成后，将pUL 25招募到门户以保留门户的功能。 病毒基因组我们将在两个具体的目标中检验假设，这两个目标一起调查 门静脉顶点的结构和功能。目的1关注的是病毒蛋白所需的 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物 反应目标2遵循一种互补的结构方法来理解 入口顶点，包括不同装配阶段所需的组件， 包装.我们的研究结果将为创新和特定疗法的发展提供信息 旨在通过阻断DNA包装来防止疱疹病毒复制。