Bridging KSHV capsids to the nuclear egress complex

将 KSHV 衣壳桥接至核出口复合体

• 批准号: 8570507
• 负责人: PRASHANT J DESAI
• 金额: $ 22.84万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-07 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8570507
• 关键词: Antiviral Agents; Baculoviruses; Binding; Biochemical; Biological; Capsid; Capsid Proteins; Cell Nucleus; Cell membrane; Cells; Cellular Membrane; Complex; Constitution; Cytosol; DNA Viruses; Development; Electrons; Elements; Genetic; Genome; Glycoproteins; Goals; Golgi Apparatus; Herpesviridae; Human Herpesvirus 8; Immunocompromised Host; In Vitro; Insecta; Light; Link; Membrane; Methods; Modeling; Nuclear; Nuclear Envelope; Nuclear Lamina; Open Reading Frames; Outcome; Pathway interactions; Play; Protein Binding; Proteins; Recombinants; Role; Structure; System; Testing; Time; Translating; Vesicle; Viral; Virion; Virus; Virus Assembly; Virus-like particle; base; imaging modality; innovation; novel; novel vaccines; particle; pathogen; protein complex; protein expression; public health relevance; scaffold; self assembly; tumor

DESCRIPTION (provided by applicant): The herpesvirus virion is comprised of three structural components: an icosahedral capsid, which encloses the genome; an electron dense asymmetrically distributed material, which immediately surrounds the capsid and is termed the tegument; and an outer membrane or envelope, which encloses the tegument and capsid and in which are embedded the viral glycoproteins. These virions are the most complex, highly organized structural entities comprised of several protein sub-units. Two studies on Kaposi's sarcoma-associated herpesvirus (KSHV) from our lab have accomplished the self-assembly of icosahedral capsids and re-constitution of the nuclear egress complex (NEC), in insect cells using baculoviruses for protein expression. This has given us a unique opportunity and the impetus to do something novel and innovative that could have the potential to be translated into the development of virus-like particles (VLPs) for new vaccine platforms. The aims will focus on beginning to physically link KSHV capsids with the NEC using biochemical and cell biological methods in this "ex-vivo" culture system. We will test capsid-associated proteins, for example, the capsid vertex-specific component (CVSC) encoded by open reading frames (ORF) 19 and 32 for the ability to bridge capsids with nuclear membranes. The outcomes will also shed light on the mechanism of capsid maturation for KSHV and for other herpesviruses. Specific Aim 1. Elucidate the interactions and capsid association of the KSHV CVSC. (a). Interactions of ORF32 and ORF19 with the capsid proteins. (b). Binding of the CVSC to KSHV icosahedral capsids. Specific Aim 2. Discover how to physically link KSHV capsids with the NEC. (a). Interactions of the NEC with the CVSC. (b). Link assembled capsids with the NEC in nuclear membranes. This proposal will discover if the "ex-vivo" produced KSHV capsids can be "decorated" by the CVSC and then test these for their potential to bind to the nuclear egress complex. The final goal of this proposal is to link these structural entities together. The outcome of this approach if successful is the use of these particles for new vaccine platforms for a clinically important pathogen and to develop new antiviral strategies.

描述（由申请人提供）：疱疹病毒病毒粒子由三种结构成分组成：二十面体衣壳，包裹基因组；一种电子密集的不对称分布的物质，直接包裹在衣壳周围，被称为衣壳；外膜或包膜，包裹着被皮和衣壳，里面嵌入了病毒糖蛋白。这些病毒粒子是由几个蛋白质亚基组成的最复杂、高度组织化的结构实体。本实验室对卡波西肉瘤相关疱疹病毒（KSHV）的两项研究，利用杆状病毒在昆虫细胞中表达蛋白质，完成了二十面体衣壳的自组装和核出口复合体（NEC）的重建。这给了我们一个独特的机会和动力去做一些新颖和创新的事情，这些事情有可能转化为开发用于新疫苗平台的病毒样颗粒（vlp）。目标将集中在开始使用生化和细胞生物学方法在这种“离体”培养系统中将KSHV衣壳与NEC物理连接起来。我们将测试衣壳相关蛋白，例如，由开放阅读框（ORF） 19和32编码的衣壳顶点特异性成分（CVSC），以测试其连接衣壳和核膜的能力。这些结果也将揭示KSHV和其他疱疹病毒衣壳成熟的机制。具体目标阐明KSHV CVSC的相互作用和衣壳关联。(一)。ORF32和ORF19与衣壳蛋白的相互作用。(b)。CVSC与KSHV二十面体衣壳的结合。具体目标2。了解如何物理连接KSHV衣壳与NEC。(一)。NEC与CVSC的相互作用。(b)。林克用核膜上的NEC组装衣壳。该方案将发现“离体”产生的KSHV衣壳是否可以被CVSC“修饰”，然后测试它们与核出口复合物结合的潜力。本提案的最终目标是将这些结构实体联系在一起。如果成功，这种方法的结果是将这些颗粒用于临床重要病原体的新疫苗平台并开发新的抗病毒策略。