Epstein-Barr virus glycoproteins and virus spread

EB 病毒糖蛋白和病毒传播

• 批准号: 7557821
• 负责人: Lindsey M. Hutt-Fletcher
• 金额: $ 30.35万
• 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 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7557821
• 关键词: Address; Adult; B-Lymphocytes; Behavior; Binding Sites; Biochemical; Burkitt Lymphoma; Capsid; Cell Nucleus; Cell fusion; Cells; Complex; Cytoplasm; Cytoplasmic Tail; Defect; Development; Disease; Dissociation; Epstein-Barr Virus latency; Extracellular Domain; Family member; Glycoproteins; Goals; Herpesviridae; Hodgkin Disease; Human Herpesvirus 4; Individual; Infection; Laboratories; Large-Cell Immunoblastic Lymphoma; Link; Maps; Membrane Proteins; Movement; Mutate; Mutation; Nasopharynx Carcinoma; Nuclear Envelope; Nuclear Outer Membrane; One-Step dentin bonding system; Penetration; Phenotype; Play; Population; Process; Production; Proteins; Research; Risk Factors; Role; Serological; Staging; Structural Protein; Testing; Tropism; Viral Proteins; Viral load measurement; Virion; Virus; Virus Assembly; Virus Replication; Work; Yeasts; env Gene Products; lytic replication; malignant stomach neoplasm; meetings; novel; protein function; recombinant virus; tool; tumor; 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）：全球 95% 以上的成年人携带 Epstein-Barr 病毒 (EBV)，与免疫母细胞淋巴瘤、伯基特淋巴瘤、霍奇金病、鼻咽癌和胃癌的发生有关。 疾病的最直接原因是潜伏感染细胞的行为。 然而，高效复制对于病毒在宿主内部和宿主之间的传播至关重要，并会影响病毒载量。 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。 这项研究的长期目标是了解 EBV 膜蛋白在向性和复制效率中发挥的作用。 该应用的重点是了解糖蛋白 gB 和 gNgM 以及包膜蛋白 BFRF1 的多种功能以及它们在病毒进入或传播中发挥的作用。 糖蛋白 gB 参与病毒组装，对病毒细胞融合很重要；缺乏 gN 的病毒无法表达 gM，并且在组装和渗透过程中存在衣壳和包膜的缔合缺陷以及可能的解离缺陷。 BFRF1 与 BFLF2 相互作用，可能在退出细胞核中发挥作用。 酵母双杂交筛选揭示了 gB 细胞质尾部与已知或被认为参与病毒组装的蛋白质之间的相互作用。 目标 1 将通过确定这些相互作用是否可以通过生化方法复制、绘制 gB 上的结合位点并确定结合位点的突变是否复制缺乏 gB 的病毒表型来测试这些相互作用的重要性。 这将通过用反式表达的突变 gB 拯救 gB 负病毒或通过在新衍生的 EBV-Bac 中构建突变来完成。 目标 2 将检验 gB 在病毒粒子与初级包膜和外核膜融合中发挥作用的假设。 将检查细胞质尾部含有阻止病毒细胞融合的突变的gB，或细胞外结构域含有突变的gB，以确定其将病毒从细胞核递送至细胞质的能力。 目标 3 将检查缺乏 BFRF1 或其伙伴 BFLF2 的病毒的表型。 目标 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化分析，以寻找与 gM 长胞质尾相互作用的蛋白质，这些蛋白质可能有助于 gNgM 负病毒的表型，并且将检查缺乏 gM 胞质尾的病毒的表型。 将检查 gM 对其他糖蛋白募集的影响。

项目成果

Epstein-Barr virus glycoprotein gM can interact with the cellular protein p32 and knockdown of p32 impairs virus.

• DOI: 10.1016/j.virol.2015.12.019
• 发表时间: 2016-02
• 期刊: Virology
• 影响因子: 3.7
• 作者: Changotra H;Turk SM;Artigues A;Thakur N;Gore M;Muggeridge MI;Hutt-Fletcher LM
• 通讯作者: Hutt-Fletcher LM