Epstein-Barr virus glycoproteins and virus spread

EB 病毒糖蛋白和病毒传播

• 批准号: 6984796
• 负责人: Lindsey M. Hutt-Fletcher
• 金额: $ 31.86万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6984796
• 关键词: Epstein Barr virus; binding sites; capsid; gene mutation; glycoproteins; laboratory rabbit; laboratory rat; latent virus infection; protein protein interaction; protein structure function; virion; virus assembly; virus envelope; virus genetics; virus infection mechanism; virus protein; virus receptors; virus related neoplasm /cancer; virus replication; yeast two hybrid system

DESCRIPTION (provided by applicant): Epstein-Barr virus (EBV) is carried by more than 95% of the adult population worldwide and is causally implicated in the development of immunoblastic lymphoma, Burkitt's lymphoma, Hodgkin's Disease, nasopharyngeal carcinoma and gastric cancer. The proximal cause of disease is the behavior of a latently infected cell. However, productive replication is critical to spread of virus within and between hosts and impacts virus load. Serologic evidence of increased virus replication is a risk factor for development of some EBV-associated tumors. The long-term goal of this research is to understand the roles that the membrane proteins of EBV play in tropism and efficiency of replication. The application focuses on understanding the multiple functions of glycoproteins gB and gNgM and the envelope protein BFRF1 and the roles they play in virus entry or spread. Glycoprotein gB has been implicated in virus assembly and is important to virus cell fusion; virus lacking gN fails to express gM and has a defect in association and possibly also in dissociation of capsids and envelope during assembly and penetration. BFRF1 interacts with BFLF2 and may play a role in exit from the nucleus. A yeast two-hybrid screen has revealed interactions between the cytoplasmic tail of gB and proteins known or thought to be involved in virus assembly. Aim 1 will test the significance of these interactions by determining if they can be reproduced biochemically, mapping the binding sites on gB and determining if mutation of binding site(s) reproduces the phenotype of a virus that lacks gB. This will be done by rescuing gB minus virus with mutated gB expressed in trans or by building mutations into a newly derived EBV-Bac. Aim 2 will test the hypothesis that gB plays a role in fusion of virions with a primary envelope and the outer nuclear membrane. gB containing mutations in the cytoplasmic tail that block virus cell fusion, or with mutations in the extracellular domain will be examined for the ability to deliver virus from the nucleus to the cytoplasm. Aim 3 will examine the phenotype of a virus that lacks BFRF1 or its partner BFLF2. Aim 4 will compare the phenotype of a virus genetically lacking gM with one that lacks gM. Yeast two-hybrid and biochemical analysis will be done to look for proteins whose interactions with the long cytoplasmic tail of gM may contribute to the phenotype of a gNgM minus virus and the phenotype of a virus that lacks the cytoplasmic tail of gM will be examined. Effects of gM on recruitment of other glycoproteins will be examined.

描述（由申请人提供）：EB病毒（EBV）在全球超过95%的成年人群中携带，并与免疫母细胞性淋巴瘤、伯基特淋巴瘤、霍奇金病、鼻咽癌和胃癌的发生有关。 疾病的近因是潜伏感染细胞的行为。 然而，生产性复制对于病毒在主机内和主机之间的传播至关重要，并影响病毒负载。 病毒复制增加的血清学证据是某些EBV相关肿瘤发生的危险因素。 本研究的长期目标是了解EB病毒膜蛋白在嗜性和复制效率中的作用。 该应用程序的重点是了解糖蛋白gB和gNgM和包膜蛋白BFRF 1的多种功能，以及它们在病毒进入或传播中所起的作用。 糖蛋白gB与病毒组装有关，对病毒细胞融合很重要;缺乏gN的病毒不能表达gM，在组装和穿透过程中，衣壳和包膜的结合以及可能的解离存在缺陷。 BFRF 1与BFLF 2相互作用，并可能在退出细胞核中发挥作用。 酵母双杂交筛选揭示了gB的胞质尾区与已知或被认为参与病毒组装的蛋白质之间的相互作用。 目的1将通过确定这些相互作用是否可以生物化学地再现、绘制gB上的结合位点并确定结合位点的突变是否再现缺乏gB的病毒的表型来测试这些相互作用的意义。 这将通过用反式表达的突变gB拯救gB减病毒或通过将突变构建到新衍生的EBV-Bac中来完成。 目的2将检验gB在病毒体与初级包膜和外核膜的融合中起作用的假设。 将检查胞质尾区中含有阻断病毒细胞融合的突变或胞外结构域中具有突变的gB将病毒从细胞核递送至细胞质的能力。 目的3将检查缺乏BFRF 1或其配偶体BFLF 2的病毒的表型。 目的4将比较基因上缺乏gM的病毒与缺乏gM的病毒的表型。 将进行酵母双杂交和生物化学分析，以寻找与gM的长胞质尾相互作用可能有助于gNgM阴性病毒表型的蛋白质，并将检查缺乏gM胞质尾的病毒表型。 将检查gM对其他糖蛋白募集的影响。