PROTEIN INTERACTIONS IN HERPESVIRUS CELL FUSION

疱疹病毒细胞融合中的蛋白质相互作用

• 批准号: 2671615
• 负责人: Robert James Geraghty
• 金额: $ 3.05万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2671615
• 关键词: PROTEIN; INTERACTIONS; HERPESVIRUS; CELL; FUSION

The long term objective of this research proposal is to identify the viral and cellular components involved in cell fusion mediated by herpes simplex virus (HSV). An intimate understanding of cell fusion induced by HSV may facilitate effective treatments to prevent such cell-to-cell spread in infected individuals. An understanding of cell-to-cell fusion mediated by HSV may also provide insight into virus-to-cell fusion because the two processes require overlapping sets of proteins. This proposal outlines three different approaches to identify cellular or viral proteins that interact with the mutant or wild-type forms of two envelope glycoproteins of HSV-1 required for cell fusion, gB and gH. The first approach will be to utilize the two-hybrid system in yeast to identify gB- or gH-interacting proteins. The two-hybrid system is a genetic screen performed in yeast expressing a cDNA library to discover viral or cellular proteins that interact with mutant or wild-type cytoplasmic domains of gB or gH. The second approach will be to attempt to co-immunoprecipitate cellular or viral proteins that are interacting with mutant or wild-type forms of gB or gH in infected cells. The third approach will be to fuse mutant or wild- type versions of the cytoplasm tail of gB or gH to glutathione-S- transferase and determine if cellular or viral proteins will bind to such fusion proteins that are immobilized on columns.

该研究建议的长期目标是确定 参与细胞融合介导的病毒和细胞成分 由单纯疱疹病毒（HSV）。 对细胞的亲密理解 HSV诱导的融合可能有助于有效治疗 防止这种细胞间传播在受感染的个体中。一个 了解由HSV介导的细胞到细胞融合 提供有关病毒至细胞融合的洞察力，因为这两个过程 需要重叠的蛋白质集。 该提议概述了三个 鉴定细胞或病毒蛋白的不同方法 与两个信封的突变体或野生型形式相互作用 细胞融合，GB和GH所需的HSV-1的糖蛋白。这 第一种方法将是利用酵母中的两个杂交系统 识别GB-或GH相互作用蛋白。两个杂交系统是 在酵母中表达cDNA库的遗传屏幕 发现与突变体相互作用的病毒或细胞蛋白 GB或GH的野生型细胞质结构域。第二种方法将 试图共免疫沉淀细胞或病毒蛋白 与突变体或野生型GB或GH相互作用 感染细胞。第三种方法是融合突变体或野生 GB或GH的细胞质尾巴的类型版本到谷胱甘肽-S- 转移酶并确定细胞或病毒蛋白是否会结合 对于固定在色谱柱上的这种融合蛋白。