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顶点与 复制病毒DNA和HSV包装机器。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。为了 1 型单纯疱疹病毒 (HSV-1)，这个过程需要七个病毒基因的产物： UL6、UL15、UL17、UL25、UL28、UL32 和 UL33。大多数蛋白质参与衣壳组装 DNA 包装是保守的，这表明这些机制对于所有 疱疹病毒。 Homa 和 [CJF1] 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，用于确定相互作用 使用衣壳门户顶点切割/包装蛋白质，从而实现单位的​​稳定包装 长度基因组。了解疱疹病毒门户结构及其与 由于之间细微的形态差异，包装机械的发展受到了阻碍 门户和五邻体顶点。然而，冷冻电镜技术的最新进展使得 将门户定位在完整衣壳的图像中，产生重要的结构信息 门户网站和相关蛋白质。 pUL6 门户十二聚体似乎锚定在 衣壳通过与周围三链体的相互作用以及归因于的螺旋密度 衣壳顶点特异性成分 (CVSC) 的 pUL17 和 pUL25 亚基。在这里我们建议 结合生化和结构研究，重点关注野生型独特的门静脉顶点 HSV 和包装突变体将提供对 HSV 基因组机制的深入了解 包装。我们的目标是了解 CVSC 组件的功能及其与 门顶点和终止酶复合物切割和保留包装的病毒 基因组。根据已发表的研究和初步研究，我们相信 pUL17 正在成为关键 衣壳组装和成熟的参与者。我们假设 pUL17 锚定终止酶 复杂到门户，并在包装​​完成后，将 pUL25 招募到门户以保留 病毒基因组。我们将在两个具体目标中检验该假设，共同研究 门户顶点的结构和功能。目标 1 侧重于所需的病毒蛋白 门户复合体与复制的病毒基因组相互作用并启动切割包装 反应。目标 2 遵循互补的结构方法来理解 门户顶点，包括不同组装阶段所需的组件和 包装。我们的结果将为创新和特异性疗法的开发提供信息 旨在通过阻断 DNA 包装来防止疱疹病毒复制。