EBV gB Function in Viral Fusion, Entry, and Egress

EBV gB 在病毒融合、进入和排出中的功能

• 批准号: 7797000
• 负责人: Richard M Longnecker
• 金额: $ 4.07万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-15 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7797000
• 关键词: AIDS-Related Opportunistic Infections; Acquired Immunodeficiency Syndrome; Antibodies; B-Lymphocytes; Binding; Biological Assay; Capsid; Cell Surface Receptors; Cell fusion; Cell surface; Cells; Chimeric Proteins; Complement 3d Receptors; Cytoplasmic Tail; Cytoskeleton; Disease; EBV-associated disease; Epithelial; Epithelial Cells; Epithelial Receptor Cell; Epstein-Barr Virus Infections; Expression Library; Family; Glycoproteins; Goals; Golgi Apparatus; Guanine; Guanine Nucleotide Exchange Factors; Herpesviridae; Human; Human Herpesvirus 4; Immune; Immunoprecipitation; Infection; Knowledge; Laboratories; Lymphoid; Maintenance; Malignant - descriptor; Malignant Neoplasms; Mediating; Molecular; Monitor; Monoclonal Antibodies; Morphogenesis; Mutation; Patients; Peripheral; Phenotype; Predisposition; Process; Property; Protein Binding; Proteins; Recombinants; Regulation; Research; Role; Screening procedure; Site; Specificity; Tail; Tissues; Tropism; Viral; Viral Proteins; Virion; Virus; Virus-Induced Membrane Fusion; Work; Yeasts; base; cDNA Expression; cell type; in vivo; insight; mutant; novel; ras-Related G-Proteins; receptor; receptor function; research study; therapeutic development; tissue tropism; tool; trafficking; yeast two hybrid system

The understanding of the molecular basis of Epstein-Barr virus (EBV)entry into target cells and virion trafficking in infected cells is the long-term goal of the Lonanecker Laboratory. We anticipate that discoveries related to EBV entry and virion morphogenesis in infected cells will be important for the development of therapeutics to treat EBV-associated cancers in the human host. Our overall hypothesis that drives our research focus is that EBV gB interacts with specific cell surface receptors that facilitate viral fusion and entry into EBV target cells. In addition, the cytoplasmic tail of EBV gB interacts with host and viral proteins and this interaction is important for regulating fusion, but also required for proper egress of EBV capsids from infected cells. The tissue tropism for EBV in vivo is largely limited to cells of epithelial or lymphoid origin. The cellular and viral factors required for EBV entry of target B cells has been fairly well described. The major viral envelope glycoprotein 350/220 (gp350/220) binds to CR2/CD21 that is abundantly expressed on B cells. Subsequently, gp42 binds to HLA Class II triggering fusion mediated by gB and gH/gL. Few details are known in regard to the mechanism of EBV induced membrane fusion and viral entry into epithelial cells. It is apparent that other receptors function in epithelial cells since CD21 and HLA Class II are not typically expressed on epithelial cells. Lindsey Hutt-Fletcher has provided compelling evidence to suggest that EBV entry of epithelial cells when compared to B cells is mechanistically different. In our preliminary studies, we have shown that in contrast to B cells, which require gp42, gB, and gH/gL for efficient cell fusion, epithelial cells require only gB, and gH/gL and when a mutant form of gB is used, only gB is required for efficient cell fusion indicating that gB may be the major fusion protein for epithelial cells and that a specific epithelial receptor for gB may exist. This proposal will analyze: 1 - The role of gB in EBV entry of epithelial and B cells by the identification of important gB functional domains by site-specific and random mutation. 2 - The role of the gB cytoplasmic tail in regulating fusion and mediating virion morphogenesis as well as the function of several cellular proteins that bind the gB tail will be determined. 3 - The existence of a novel gB receptor will be investigated. Clarifying the interactions between EBV and target cells is essential for understanding the tropism of EBV infections in the human host. Knowledge of the mechanism and viral and cellular factors required for EBV entry and replication in epithelial and B cells will provide insight into host susceptibility and will allow for the development of therapeutics for the treatment or eradication of EBV-associated diseases.

EB病毒进入靶细胞和病毒粒子的分子基础 贩运感染细胞是Lonanecker实验室的长期目标。我们预计， 与感染细胞中EBV进入和病毒体形态发生相关的基因的表达对于 本发明提供了用于治疗人类宿主中EBV相关癌症的治疗剂。我们的总体假设， 研究重点是EB病毒gB与特定的细胞表面受体相互作用，促进病毒融合， 进入EBV靶细胞。此外，EBV gB的胞质尾区与宿主和病毒蛋白相互作用 这种相互作用对于调节融合是重要的，但也是EBV衣壳从细胞中正确排出所必需的。 被感染的细胞EBV在体内的组织嗜性主要限于上皮或淋巴起源的细胞。 EBV进入靶B细胞所需的细胞和病毒因子已经有相当好的描述。的 主要病毒包膜糖蛋白350/220（gp 350/220）与CR2/CD 21结合，CR2/CD 21在 B细胞。随后，gp 42与HLA II类结合，触发由gB和gH/gL介导的融合。一些细节 关于EBV诱导的膜融合和病毒进入上皮细胞的机制是已知的。 很明显，其他受体在上皮细胞中起作用，因为CD 21和HLA II类不是典型的 在上皮细胞上表达。Lindsey Hutt-Fletcher提供了令人信服的证据表明，EBV 与B细胞相比，上皮细胞的进入在机制上是不同的。在初步研究中，我们 已经表明，与需要gp 42、gB和gH/gL进行有效细胞融合的B细胞相反，上皮细胞 细胞仅需要gB和gH/gL，并且当使用gB的突变形式时，高效细胞仅需要gB 融合表明gB可能是上皮细胞的主要融合蛋白， 可能存在gB受体。该提案将分析： 1 -通过鉴定重要的gB功能，gB在EBV进入上皮细胞和B细胞中的作用 结构域的位点特异性和随机突变。 2 -gB胞质尾区在调节融合和介导病毒体形态发生中的作用以及 将测定结合gB尾的几种细胞蛋白的功能。 3 -将研究新的gB受体的存在。 阐明EBV与靶细胞之间的相互作用对于理解EBV的向性至关重要 感染人类宿主。了解EBV的机制以及所需的病毒和细胞因子 在上皮细胞和B细胞中的进入和复制将提供对宿主易感性的深入了解，并将允许 开发用于治疗或根除EBV相关疾病的治疗剂。