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.

抽象的 该项目旨在阐明单纯疱疹病毒门户顶点与 复制的病毒DNA和HSV包装机械。包装和乳沟 复制的病毒DNA进入预制的衣壳是疱疹病毒感染期间的关键步骤。为了 单纯疱疹病毒1型（HSV-1），此过程需要七个病毒基因的产物： UL6，UL15，UL17，UL25，UL28，UL32和UL33。大多数参与衣壳组件的蛋白质 DNA包装是保守的，表明所有机制也将与所有机制相似 疱疹病毒。 HOMA和[CJF1] Conway Laboratories的拟议研究结合了 病毒学和结构生物学方面的专业知识，用于确定的相互作用 裂解/包装蛋白带有衣壳门户顶点，导致单元的稳定包装 长度基因组。了解疱疹病毒门户结构及其与 由于包装机械因形态学的细微差异而受到阻碍 门户和彭顿顶点。但是，冷冻技术的最新进展允许 在完整的衣壳图像中局部的门户，得出有关的重要结构信息 门户和相关蛋白。 PUL6 Portal DoteCamer似乎已锚定 与五体三环的相互作用以及螺旋密度归因于capsid Capsid顶点特异性分量（CVSC）的PUL17和PUL25亚基。我们在这里提出 集中的生化和结构研究，重点是野生类型的独特门户顶点 HSV和包装突变体将提供有关HSV基因组机制的见解 包装。我们旨在了解CVSC组件的功能，它们与 门户顶点和末端酶复合物在包装病毒的裂解和保留方面 基因组。根据已发表和初步研究，我们认为PUL17正在成为关键 Capsid组装和成熟的玩家。我们假设PUL17锚定终端酶 到达门户的复杂，包装完成后，将PUL25招募到门户以保留 病毒基因组。我们将以两个具体的目的测试该假设，共同研究 门户顶点的结构和功能。 AIM 1专注于该病毒蛋白 门户综合体与复制的病毒基因组相互作用并启动裂解包装 反应。 AIM 2遵循一种互补的结构方法来了解 门户顶点，包括不同阶段的组件所需的组件和 包装。我们的结果将为创新和特定治疗剂的发展提供信息 旨在通过阻止DNA包装来防止疱疹病毒复制。