RNA-ENVELOPE VIRUS FORMATION IN ANIMAL CELLS

动物细胞中 RNA 包膜病毒的形成

• 批准号: 3128838
• 负责人: MILTON J SCHLESINGER
• 金额: $ 19.35万
• 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 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3128838
• 关键词: Sindbis virus; crystallization; fatty acylation; gene expression; glycoproteins; high performance liquid chromatography; membrane fusion; membrane proteins; protein structure function; radiotracer; site directed mutagenesis; temperature sensitive mutant; tissue /cell culture; transfection; virus RNA; virus classification; virus envelope; virus genetics; virus protein; virus replication

Enveloped RNA viruses are the etiological agents for several important human diseases, including acquired immune deficiency syndrome (Human Immunodeficiency Virus), influenza (Influenza Virus), lower respiratory disease in children (Respiratory Syncytial Virus) and dengue fever (Dengue Virus). Although these viruses are distinct with regard to the structure of their RNA genome and employ different strategies in their replication in host cells, they share several activities and properties in common. One of these occurs in the final stages of virus replication when virus nucleocapsids or nucleoproteins interact with virus-specific transmembranal glycoproteins to initiate the assembly of newly replicated viruses. This end-stage of the virus replication cycle is clearly essential for virus growth and spread, yet little is known about the detailed biochemistry of enveloped virus assembly. Much of the experimental work proposed in this grant application focuses on virus assembly and utilizes a model virus system which has proved to be highly amenable to a molecular biology study of virus replication. The virus is Sindbis, an alphavirus whose complete genome of 11,700 nucleotides has been cloned as a cDNA which, upon in vitro transcription, produces a precise copy of the genome RNA that is infectious in tissue culture cells and gives rise to progeny virus. The power of this system lies in the ability to prepare site-directed mutations in those regions of the virus genome which code for proteins that function in virus assembly. Experiments performed thus far with this system have yielded mutants whose phenotypes are indicative of defects in very late stages of virus replication and affect the budding process. I propose to continue and extend these studies in order to obtain a molecular description of virus nucleocapsid interactions with glycoproteins. A new collaborative project will be initiated. Its objective is to obtain a detailed atomic structure of intact Sindbis virus derived from analyses of diffraction patterns from virus crystals. Results from these studies are expected to reveal new information about enveloped virus structure and assembly and should offer new insights into the design of effective antiviral agents.

包裹的RNA病毒是几种病因 重要的人类疾病，包括获得的免疫缺陷 综合征（人类免疫缺陷病毒），流感（流感 病毒），儿童的下呼吸道疾病（呼吸道 合成病毒）和登革热（登革热病毒）。 虽然这些 病毒在其RNA的结构方面是不同的 基因组并在复制中采用不同的策略 宿主单元，它们共享几个活动和特性 常见的。 其中之一发生在病毒的最后阶段 当病毒核苷酸或核蛋白相互作用时复制 用病毒特异性的跨膜糖蛋白启动 组装新复制的病毒。 这个终点 病毒复制周期显然对于病毒生长和 传播，但对详细的生物化学知之甚少 包裹的病毒组装。 提出的许多实验工作 在此赠款中，申请专注于病毒组装并利用 一种模型病毒系统，被证明是高度适合的 病毒复制的分子生物学研究。 病毒是 信德比斯（Sindbis），一个α的完整基因组11,700 核苷酸已被克隆为cDNA，在体外 转录，产生基因组RNA的精确副本 在组织培养细胞中感染性，并引起后代 病毒。 该系统的力量在于准备的能力 病毒基因组的那些区域的位置突变 哪些在病毒组装中起作用的蛋白质代码。 到目前为止，该系统执行的实验已经产生 表型的突变体很晚有缺陷 病毒复制的阶段并影响萌芽过程。 我 建议继续并扩展这些研究，以获得 病毒核素的分子描述与 糖蛋白。 将启动一个新的协作项目。 它的目的是获得完整的详细原子结构 来自从衍射模式的分析得出的信德比斯病毒 病毒晶体。这些研究的结果有望揭示 有关包围病毒结构和组装的新信息以及 应该为有效抗病毒设计提供新的见解 代理商。