PROPERTIES OF HSV-1 GYLCOPROTEINS AND MEMBRANE PROTEINS

HSV-1 糖蛋白和膜蛋白的特性

• 批准号: 3070909
• 负责人: THOMAS C HOLLAND
• 金额: $ 6.28万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3070909
• 关键词: Herpes simplex disease; genetic translation; glycoproteins; herpes simplex virus 1; hybridomas; immunoprecipitation; laboratory rabbit; membrane fusion; membrane proteins; molecular cloning; monoclonal antibody; plasmids; point mutation; protein engineering; protein structure; synthetic peptide; virus cytopathogenic effect; virus envelope; virus genetics; virus infection mechanism; virus protein; virus replication

Infections caused by herpes simplex virus type 1 (HSV-1) and the related viral serotype HSV-2 are among the most common human infections. One of the important features of the virus is its envelope or membrane. Among the viral proteins found in this envelope are the viral glycoproteins, gB, gC, gD, dE, gG, and gH. These glycoproteins are the first virion components to come in contact with the host cell and are involved in virion attachment and penetration. Later in virus replication they become involved in envelopment and egress. Important questions concerning the role of the glycoproteins in these processes remain unresolved. There are also a number of fundamental questions concerning basic glycoprotein properties, such as membrane anchoring, which need to be answered. HSV-1 gC is a nonessential viral glycoprotein which can be used as a model glycoprotein to answer certain questions concerning membrane anchoring. The role of the transmembrane domain has been well established. Preliminary data indicate that the cytoplasmic domain is also important, perhaps due to charged amino acid residues that cannot pass through the membrane. Oligo nucleotide directed mutagenesis will be used to create additional gC mutants affected in this domain. Also, the process by which gC becomes incorporated into the virion envelope will be studied by use of an antibody selection technique to select and identify mutants which produce gC but fail to incorporate it into the virion. HSV-1 gB has been implicated in the membrane fusion process, both in penetration and cell fusion. Structural and genetic studies have identified the cytoplasmic and transmembrane domains of gB as important in the fusion process. Oligonucleotide mutagenesis will be used to construct gB mutants designed to explore the structure of this glycoprotein and to examine its role in membrane fusion. A gene apparently encoding a nonglycosylated membrane protein has also been identified, although the protein itself has not been found, perhaps because it is extremely hydrophobic. Several methods will be used to identify the primary translation product of this gene, including the production of antibodies against synthetic peptides and in vitro translation of messenger RNA.

由 1 型单纯疱疹病毒 (HSV-1) 和 相关病毒血清型 HSV-2 是最常见的人类病毒之一 感染。 该病毒的重要特征之一是 信封或膜。 在此发现的病毒蛋白中 包膜是病毒糖蛋白、gB、gC、gD、dE、gG 和 gH。 这些糖蛋白是第一个进入的病毒体成分 与宿主细胞接触并参与病毒颗粒附着 和渗透力。 后来在病毒复制过程中它们参与其中 在包围和出口中。 有关的重要问题 糖蛋白在这些过程中的作用仍未解决。 还有一些基本问题涉及 糖蛋白的基本特性，例如膜锚定， 需要回答。 HSV-1 gC 是一种非必需病毒 糖蛋白，可以作为模型糖蛋白来回答 关于膜锚定的某些问题。 的作用 跨膜结构域已经确定。 初步数据表明，细胞质结构域也是 重要的，可能是由于带电荷的氨基酸残基 不能穿过膜。 寡核苷酸定向 诱变将用于产生额外受影响的 gC 突变体 在这个域中。 另外，gC 变成的过程 并入病毒颗粒包膜将通过使用进行研究 抗体选择技术来选择和鉴定突变体 产生 gC 但未能将其整合到病毒体中。 HSV-1gB 与膜融合过程有关 渗透和细胞融合。 结构和遗传学研究 将 gB 的细胞质和跨膜结构域鉴定为 在融合过程中很重要。 寡核苷酸诱变将 用于构建旨在探索结构的 gB 突变体 这种糖蛋白并检查其在膜融合中的作用。 显然编码非糖基化膜蛋白的基因 也已被鉴定，尽管蛋白质本身尚未被鉴定 发现，也许是因为它极其疏水。 一些 将使用方法来识别主要翻译产品 该基因的作用，包括产生针对 合成肽和信使RNA的体外翻译。