Structure and function of EBV protein complexes that trigger epithelial cell entry

触发上皮细胞进入的 EBV 蛋白复合物的结构和功能

• 批准号: 9093710
• 负责人: Theodore S Jardetzky
• 金额: $ 19.54万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9093710
• 关键词: Adopted; Affinity; Antiviral Agents; Architecture; B-Lymphocytes; Binding; Biophysics; Burkitt Lymphoma; C-terminal; Cell fusion; Cells; Cellular Membrane; Collaborations; Comparative Study; Complex; Cryoelectron Microscopy; Defect; Disease; Electron Microscopy; Epithelial; Epithelial Cells; Epstein-Barr Virus Infections; Event; Family; Glycoproteins; Goals; Grant; HLA Antigens; Health; Herpesviridae; Hodgkin Disease; Human; Human Herpesvirus 4; Individual; Infectious Mononucleosis; Integrin Binding; Integrins; Laboratories; Life; Malignant Neoplasms; Malignant lymphoid neoplasm; Mediating; Membrane; Membrane Fusion; Methods; Modeling; Molecular Conformation; Mutagenesis; Mutation; Nasopharynx Carcinoma; Negative Staining; Oncogenic Viruses; Physiological; Process; Proteins; Receptor Cell; Research; Resolution; Role; Specificity; Stomach Carcinoma; Structure; Subgroup; Viral; Viral Proteins; Virus; Virus Receptors; cell type; falls; gammaherpesvirus; in vivo; insight; member; mutant; novel strategies; programs; protein complex; receptor; receptor binding; reconstitution; tumor; vaccine development; virology

 DESCRIPTION (provided by applicant): Epstein-Barr virus (EBV) is a member of the herpes virus family, causing infectious mononucleosis and establishing life-long latency in infected individuals. EBV is also associated with cancers of both B cells and epithelial cells, the two cell types that it infects in vivo. EBV entry into B cells and epithelial cells requires the coordinated action of four (gH, gL, gB and gp42) and three (gH, gL and gB) viral glycoproteins, respectively. The process by which receptor recognition triggers membrane fusion and virus entry is not well understood. The gH, gL and gB proteins form the core fusion machinery for all herpesviruses and it is thought that gB acts as the primary fusogen in this process. The gH/gL heterodimer is thought to act as the regulator for gB activation, triggering conformational changes in gB after receptor binding. We recently determined the low- resolution electron microscopy structure of the EBV B cell entry triggering complex composed of gH/gL, gp42 and host receptor HLA, clarifying how this complex bridges the viral and cellular membranes, bringing them into closer proximity prior to gB activation. Here, we (the Jardetzky, Longnecker and Zhou research groups) propose to study complexes of wild type and mutant gH/gL with integrins that act as the entry receptor for EBV infection of epithelial cells by a combination of virology, mutagenesis and biophysics means. We will first characterize molecular interactions between the wt (wild type) or mutant gH/gL proteins and integrin receptors and determine how initial complexes between gH/gL and integrins may convert to an activated state to trigger gB- mediated fusion. We will then compare the architectures of the B cell and epithelial cell triggering complexes by cryo electron microscopy. The anticipated results would establish the molecular interactions of the two EBV entry complexes and reveal the structural commonalities between them. Such structural information is key to understanding the subsequent steps of gB activation and EBV entry and, by extension, the general mechanism of cell entry by human herpesviruses.

 描述(申请人提供)：爱泼斯坦-巴尔病毒(EBV)是疱疹病毒家族的成员，会导致传染性单核细胞增多症，并在感染者中建立终身潜伏期。EB病毒也与B细胞和上皮细胞的癌症有关，这两种细胞 它在体内感染的类型。EBV进入B细胞和上皮细胞需要协调 四种病毒糖蛋白(Gh、g1、gB和GP42)和三种病毒糖蛋白(Gh、g1和gB)的作用。受体识别触发膜融合和病毒进入的过程尚不清楚。Gh、g1和gB蛋白形成了所有疱疹病毒的核心融合机制，gB被认为在这一过程中扮演着主要的FusoGen的角色。Gh/Gl异源二聚体被认为是GB激活的调节因子，在受体结合后触发GB的构象变化。我们最近测定了由Gh/g1、GP42和宿主受体HLA组成的EBV B细胞进入触发复合体的低分辨电子显微镜结构，阐明了该复合体如何将病毒膜和细胞膜连接起来，使它们在GB激活之前更接近。在这里，我们(Jaradezky、Longnecker和周的研究小组)建议通过病毒学、突变和生物物理手段相结合的方法，研究野生型和突变型Gh/gl与整合素的复合体，作为EBV感染上皮细胞的进入受体。我们将首先描述野生型或突变型Gh/gl蛋白与整合素受体之间的分子相互作用，并确定Gh/gl与整合素之间的初始复合体如何转变为激活状态以触发GB介导的融合。然后，我们将用低温电子显微镜比较B细胞和上皮细胞触发复合体的结构。预期的结果将建立两个EBV进入复合体的分子相互作用，并揭示它们之间的结构共性。这些结构信息是理解GB激活和EBV进入的后续步骤以及人类疱疹病毒进入细胞的一般机制的关键。