RNA-ENVELOPE VIRUS FORMATION IN ANIMAL CELLS

动物细胞中 RNA 包膜病毒的形成

• 批准号: 3128841
• 负责人: MILTON J SCHLESINGER
• 金额: $ 15.47万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3128841
• 关键词: Sindbis virus; cell growth regulation; cell membrane; chemical cleavage; fatty acylation; gene expression; genetic manipulation; glycoproteins; laboratory rabbit; membrane proteins; messenger RNA; protease inhibitor; proteolysis; radiotracer; temperature sensitive mutant; tissue /cell culture; virus RNA; virus classification; virus genetics; virus protein; virus replication; Sindbis virus; cell growth regulation; cell membrane; chemical cleavage; fatty acylation; gene expression; genetic manipulation; glycoproteins; laboratory rabbit; membrane proteins; messenger RNA; protease inhibitor; proteolysis; radiotracer; temperature sensitive mutant; tissue /cell culture; virus RNA; virus classification; virus genetics; virus protein; virus replication

Enveloped RNA viruses are the etiological agents for many human diseases-ranging from yellow fever, the first human disease attributable to a virus (1902), to acquired immune deficiency syndrome, caused by the retrovirus HTLVIII/LAV (1984). Although these two viruses, as well as other enveloped RNA viruses, differ importantly in their genomic structure, composition and mechanisms for gene expression, they have some activities in common in their replication cycle and all synthesize virus-specific transmembranal glycoproteins that interact with virus nucleoprotein structures during virus assembly. Experiments in this proposal are focused on this common activity but employ a simple "model" enveloped RNA virus-cell system, Sindbis virus replication in tissue culture cells. These projects constitute an ongoing research program whose goal is the elucidation of critical biochemical events in glycoprotein processing and assembly of enveloped RNA viruses. The work proposed here will utilize recombinant DNA technology to obtain site-specific mutations in a specific region of this virus' glycoprotein to determine what factors are required to achieve the highly selective interaction between the transmembranal virus "spike" proteins and nucleocapsid of the virus. Systems to measure binding between nucleocapsids and glycoproteins will also be sought. Of particular interest is what role fatty acylation, a modification present in many virus and cell glycoproteins, may play in virus assembly. In vitro mutagenesis will be used further to analyze a small region in the Sindbis genome required for processing and insertion of glycoprotein into the cell's membrane. Our ability to detect effects of directed glycoprotein changes on virus assembly are now feasible because a cDNA capable of transcription into an infectious RNA has been developed.

包裹的RNA病毒是许多人类的病因 疾病来自黄热病，第一种人类疾病 归因于病毒（1902），可获得免疫缺陷 综合征，由逆转录病毒HTLVIII/LAV（1984）引起。 虽然 这两种病毒以及其他包膜的RNA病毒都不同 重要的是它们的基因组结构，组成和 基因表达的机制，它们有一些活动 在其复制周期中常见和所有合成病毒特异性 与病毒相互作用的跨膜糖蛋白 病毒组装过程中的核蛋白结构。 实验 该提案的重点是这项共同活动，但采用了 简单的“模型”包裹的RNA病毒细胞系统，sindbis病毒 在组织培养细胞中复制。 这些项目构成 正在进行的研究计划，其目标是阐明关键 糖蛋白加工和组装中的生化事件 包裹的RNA病毒。 这里提出的工作将利用 重组DNA技术以获得位点特异性突变 该病毒糖蛋白的特定区域，以确定什么 需要因素才能实现高度选择性的互动 在跨膜病毒“尖峰”蛋白和 病毒的核包囊。 测量结合之间的系统 还将寻求核蛋白质和糖蛋白。 特别 兴趣是哪种角色脂肪酰化，这是一种修改 许多病毒和细胞糖蛋白可能会在病毒组装中发挥作用。 在 体外诱变将进一步分析 加工和插入所需的信德比斯基因组 糖蛋白进入细胞的膜。 我们检测的能力 定向糖蛋白变化对病毒组装的影响是 现在是可行的，因为能够转录到一个 已经开发了感染性RNA。