FUNCTIONAL DOMAINS OF HSV-1 GLYCOPROTEINS GB AND GC

HSV-1 糖蛋白 GB 和 GC 的功能域

• 批准号: 2061729
• 负责人: THOMAS C HOLLAND
• 金额: $ 22.57万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1996-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2061729
• 关键词: Alphaherpesvirinae; Herpes simplex disease; antibody neutralization test; cell fusion; cell membrane; gene expression; genetic mapping; genetic promoter element; genetic transcription; glycoproteins; laboratory mouse; laboratory rabbit; membrane proteins; molecular cloning; mutant; plasmids; protein biosynthesis; protein sequence; protein structure function; tissue /cell culture; virion; virus DNA; virus genetics; virus protein; virus replication

Herpes Simplex Virus Type 1 (HSV-1) is the cause of primary and recurrent infections of the mouth, gums, and lip, genital and ocular infections, generalized infections in neonates and immunodeficient individuals, and rare cases of encephalitis. The closely related virus HSV-2 is the cause of genital herpetic lesions and numerous neonatal infections. The process by which HSV-1 absorbs to and enters cells is not well understood. An understanding of this process may well lead to the development of effective and practical antiviral agents and/or immunization methods that block these early stages of infection. Several HSV-1 glycoproteins (gB, gC, gD, and gH) have been proven or are likely to be involved in these processes. Initial interaction of HSV-1 virions with the cell surface may be mediated by binding of viral glycoproteins to cell surface heparan sulfate. The identity of the viral adsorption protein that interacts with heparan sulfate is unclear, since virions lacking gB, gC, or gD all adsorb to cells. However, solubilized gB and gC bind to heparin. It has been suggested that initial binding to cells may be mediated by either glycoprotein. This hypothesis will be tested directly by the production of virions lacking both gB and gC and their use in adsorption experiments. Virions deficit in other combinations of glycoproteins will be tested as necessary. These virions will be produced by introduction of sites recognized by Factor Xa, a highly specific protease, or by recombination of existing glycoprotein deficient mutants. Once adsorbed, virions penetrate by fusion of the virion envelope with the plasma membrane. Three glycoproteins, gB, gC, and gH, have been shown to be essential for this process, although the exact roles of each are unknown. The penetration process is likely to be related to the induction of cell fusion by syncytial mutants of HSV-1, since many of the same glycoproteins have been implicated in each. Also involved in cell fusion is the UL53/syn1 gene. We have performed an extensive in vitro analysis on the protein specific by this gene. Significantly, we have found that the processed from of this protein has a hydrophobic N-terminal end, similar to the hydrophobic N- terminal fusion domains of other viral fusion proteins. It is likely that UL53 acts as a membrane fusogen in the cell fusion process and possibly in penetration as well. Extensive additional characterization of UL53 is proposed to augment our understanding of this molecule. These studies will include determination of the transmembrane structure of the UL53 protein, use of UL53 specific antisera to characterize the protein in vivo, sequencing of additional UL53 syncytial mutants, and production of new UL53 mutants. Finally, a transfection system for induction of syncytium formation by HSV-1 will be developed and used to identify the viral genes that are necessary and sufficient for cell fusion. This will eventually make it possible to probe the mechanism of cell fusion and determine its relationship to virion penetration.

单纯疱疹病毒1型（HSV-1）是主要和经常性的原因 口腔，牙龈和唇，生殖器和眼部感染的感染， 新生儿和免疫缺陷个体中的普遍感染，以及 极少数脑炎病例。 密切相关的病毒HSV-2是原因 生殖器疱疹病变和许多新生儿感染。 过程 HSV-1吸收并进入细胞尚不清楚。 一个 了解这一过程很可能导致有效的发展 和实用的抗病毒药物和/或免疫方法阻止这些方法 感染的早期阶段。 几种HSV-1糖蛋白（GB，GC，GD和 GH）已被证明或可能参与这些过程。 HSV-1病毒体与细胞表面的初始相互作用可以介导 通过将病毒糖蛋白与细胞表面硫酸肝素结合结合。 这 与乙酰肝素相互作用的病毒吸附蛋白的身份 硫酸盐尚不清楚，因为缺乏GB，GC或GD的病毒体所有吸附 细胞。 但是，溶解的GB和GC与肝素结合。 它一直 建议最初与细胞的结合可以由任何一个介导 糖蛋白。 该假设将通过生产直接检验 缺乏GB和GC的病毒体及其在吸附实验中的使用。 糖蛋白的其他组合中的病毒体赤字将被测试为 必要的。 这些病毒体将通过引入站点产生 通过因子Xa识别，高度特异性的蛋白酶，或通过重组 现有的糖蛋白缺乏突变体。 吸附后，病毒体穿透 通过融合病毒膜与质膜的融合。 三 糖蛋白，GB，GC和GH已被证明对此至关重要 过程，尽管每个的确切作用都是未知的。 穿透 过程可能与通过 HSV-1的合成突变体，因为许多相同的糖蛋白已经 与每个有关。 细胞融合也涉及的是UL53/Syn1基因。 我们已经对特有的蛋白质进行了广泛的体外分析 这个基因。 值得注意的是，我们发现从此处理的处理 蛋白质具有疏水N末端，类似于疏水N- 其他病毒融合蛋白的末端融合结构域。 很可能 UL53在细胞融合过程中充当膜融合原，可能在 渗透。 UL53的广泛表征是 提议增强我们对该分子的理解。 这些研究会 包括确定UL53蛋白的跨膜结构， 使用UL53特异性抗血清来表征体内蛋白质的表征 额外的UL53合成突变体的测序和新的UL53的产生 突变体。 最后，转染系统诱导合胞体 HSV-1的形成将开发并用于识别病毒基因 对于细胞融合而言，这是必要且足够的。 最终会 使探测细胞融合的机理并确定其可能 与病毒体渗透的关系。