PROTEIN INTERACTIONS IN HERPESVIRUS CELL FUSION

这项研究计划的长远目标是找出 参与细胞融合介导的病毒和细胞成分 单纯疱疹病毒（HSV） 深入了解细胞 HSV诱导的融合可能有助于有效治疗， 防止感染个体中的这种细胞间传播。一个 对HSV介导的细胞间融合的理解也可能 提供了对病毒与细胞融合的深入了解， 需要重叠的蛋白质 该提案概述了三个 不同的方法来鉴定细胞或病毒蛋白， 与两个包膜的突变或野生型形式相互作用 细胞融合所需的HSV-1糖蛋白，gB和gH。的 第一种方法是利用酵母中的双杂交系统， 鉴定gB或gH相互作用蛋白。双混合动力系统是 在表达cDNA文库的酵母中进行遗传筛选， 发现病毒或细胞蛋白质与突变体或 gB或gH的野生型胞质结构域。第二种方法将 试图使细胞或病毒蛋白质 与gB或gH的突变体或野生型相互作用， 被感染的细胞第三种方法是融合突变体或野生型- 型版本的gB或gH的细胞质尾谷胱甘肽-S- 转移酶，并确定细胞或病毒蛋白质是否会结合 涉及固定在柱上的这种融合蛋白。

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.