Nucleocapsid envelopment Herpes simplex virus-1

核衣壳包膜单纯疱疹病毒1

• 批准号: 7020025
• 负责人: JOEL D. BAINES
• 金额: $ 34.18万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7020025
• 关键词: electron microscopy; herpes simplex virus 1; immunoelectron microscopy; membrane structure; nuclear membrane; nucleocapsid; protein localization; virion; virus assembly; virus genetics; virus protein; virus replication

DESCRIPTION (provided by applicant): The long-term goal of these studies is to understand the molecular mechanisms of herpesvirus nucleocapsid envelopment. All herpesviruses bud initially from the inner nuclear membranes of infected cells suggesting that information gained from these studies will be applicable to all members of this medically important virus family. The application focuses on the role of herpes simplex virus 1 UL31 and UL34 in the envelopment process. Hypotheses based on extensive preliminary data are tested. The first hypothesis proposes that the protein encoded by UL31 (pUL3 t) interacts with the UL34 gene product (pUL34) to mediate proper targeting of both proteins to the inner nuclear membrane. To assess the importance of the interaction in vivo, viral mutants bearing UL31 genes that lack pUL34 interacting domains, and UL34 mutants that lack pUL31 interacting regions will be tested for their abilities to correctly target both proteins in infected cells and to mediate nucleocapsid envelopment. The second hypothesis to be tested seeks to understand the role of nuclear lamina association of pUL31. The nuclear lamina is part of the nucleocytoskeleton or nuclear matrix that lines the inside surface of the nuclear rim. One possible role is that nuclear lamina association of pUL31 mediates anchoring of the pUL31/pUL34 complex at the nuclear rim. Therefore UL31 mutants lacking lamina association domains and containing known lamina binding domains replacing these domains will be tested for their abilities to mediate correct targeting of pUL31 and pUL34 to the nuclear rim in transient expression assays. Mutant viruses expressing these proteins will then be assessed for their ability to produce enveloped virions by electron microscopic examination of cells infected with these viruses. A second possible role is that pUL31 causes partial lamina depolymerization to allow nucleocapsids access to the inner nuclear membrane. To assess this possibility, the lamina of cells infected with wild type virus and a deletion virus lacking UL31 will be characterized by immunoelectron microsopy. If the wild type virus infected cells contain a porous lamina whereas cells infected with the UL31 mutant do not, mechanisms by which pUL31 could mediate local lamina depolymerization will be explored.

描述（由申请人提供）：这些研究的长期目标是了解疱疹病毒核衣壳包膜的分子机制。所有疱疹病毒最初都是从受感染细胞的内核膜中出芽，这表明从这些研究中获得的信息将适用于这个医学上重要的病毒家族的所有成员。该应用重点关注单纯疱疹病毒 1 UL31 和 UL34 在包膜过程中的作用。基于大量初步数据的假设得到了检验。第一个假设提出，UL31 (pUL3 t) 编码的蛋白质与 UL34 基因产物 (pUL34) 相互作用，介导两种蛋白质正确靶向内核膜。为了评估体内相互作用的重要性，将测试携带缺乏 pUL34 相互作用结构域的 UL31 基因的病毒突变体和缺乏 pUL31 相互作用区域的 UL34 突变体，以确定它们正确靶向受感染细胞中的两种蛋白质和介导核衣壳包封的能力。要测试的第二个假设旨在了解 pUL31 核纤层联合的作用。核纤层是核细胞骨架或核基质的一部分，排列在核边缘的内表面上。一种可能的作用是 pUL31 的核层联合介导 pUL31/pUL34 复合物在核边缘的锚定。因此，将测试缺乏核纤层关联结构域并含有替代这些结构域的已知核纤层结合结构域的 UL31 突变体在瞬时表达测定中介导 pUL31 和 pUL34 正确靶向核边缘的能力。然后，通过电子显微镜检查感染这些病毒的细胞，评估表达这些蛋白质的突变病毒产生有包膜病毒颗粒的能力。第二个可能的作用是 pUL31 引起部分核纤层解聚，使核衣壳能够进入内核膜。为了评估这种可能性，将通过免疫电子显微镜对感染野生型病毒和缺乏 UL31 的缺失病毒的细胞层进行表征。如果野生型病毒感染的细胞含有多孔层，而感染 UL31 突变体的细胞则不含有，则将探索 pUL31 介导局部层层解聚的机制。