Biophysical and structural analysis of the herpesviral nuclear budding machinery

疱疹病毒核出芽机制的生物物理和结构分析

• 批准号: 10159089
• 负责人: Ekaterina Heldwein
• 金额: $ 59.58万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-05 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10159089
• 关键词: Affect; Animals; Antiviral Agents; Binding; Biochemistry; Biological; Biological Models; Biology; Biophysics; Blindness; Capsid; Capsid Proteins; Cell Nucleus; Cells; Chemicals; Complex; Cryoelectron Microscopy; Cytoplasm; Data; Dissection; Electron Spin Resonance Spectroscopy; Electrostatics; Encephalitis; Goals; Herpes Labialis; Herpesviridae; Human; Immunocompromised Host; In Vitro; Individual; Infection; Iowa; Knowledge; Lead; Life; Lipids; Malignant Neoplasms; Mediating; Membrane; Membrane Proteins; Modeling; Nature; Newborn Infant; Nuclear; Nuclear Envelope; Nuclear Inner Membrane; Nuclear Outer Membrane; Nucleocapsid; Phenotype; Phosphorylation; Population; Positioning Attribute; Process; Proteins; Regulation; Research; Role; Scaffolding Protein; Structure; Suppressor Mutations; System; Therapeutic; Therapeutic Intervention; Vertebrate Viruses; Vesicle; Viral; Viral Proteins; Virion; Virus; Virus Replication; Work; base; biophysical techniques; burden of illness; combat; design; driving force; ds-DNA; genital herpes; in vitro Model; innovation; interdisciplinary approach; latent infection; mutant; new therapeutic target; novel; novel strategies; novel therapeutic intervention; particle; pathogen; structural biology; virus envelope

PROJECT SUMMARY/ABSTRACT Herpesviruses are double-stranded-DNA enveloped viruses that are among the most complex viruses infecting animals. This proposal focuses on nuclear egress, a critical, conserved step in the assembly and release of progeny virions during which nucleocapsids are translocated from the nucleus into the cytoplasm where they mature into infectious virions. The viral nuclear egress complex (NEC) is the key player in this process. Using in vitro model systems, we previously discovered that the NEC is a complete, virally encoded membrane budding machine that operates at the nuclear envelope. However, a major barrier to understanding nuclear egress is the lack of knowledge of how the NEC generates membrane curvature that results in budding. The long-term goal of this research is to elucidate the detailed mechanism of herpesvirus nuclear egress, both to gain a fundamental knowledge of this unusual process and to identify and characterize novel targets for antiviral therapeutic design. This proposal is driven by the central hypothesis, based on substantial preliminary data, that both NEC/membrane interactions and NEC oligomerization into a coat are the major driving forces that enable negative membrane curvature formation and budding. The objective of this proposal is to systematically dissect the NEC budding mechanism in Herpes Simples virus (HSV) by characterizing essential protein/protein and protein/membrane interactions and budding intermediates by employing a multidisciplinary approach, which includes the cutting-edge approaches of cryoelectron microscopy and electron spin resonance. The scientific premise of the proposed work is that a comprehensive dissection of the NEC-mediated formation of negative membrane curvature is essential for unraveling the unusual mechanism of herpesviral nuclear egress and developing strategies to block it. Beyond viruses, this study will expand our limited mechanistic understanding of the mechanisms of membrane deformation in general. The proposal is innovative because it investigates an unusual mechanism, is guided by an original hypothesis, and employs novel approaches. The proposal is significant because it aims to advance our mechanistic understanding of an essential step in viral replication cycle with the goal of identifying new targets for therapeutic interventions and because it provides an opportunity to develop models of negative curvature formation, currently a black box.

项目摘要/摘要 疱疹病毒是双链DNA包膜病毒，是最复杂的感染病毒之一 动物。该提议着重于核出口，这是组装和释放的关键，保守的步骤 后代病毒体从核中将核从核转移到细胞质中 成熟成传染病。病毒核出口络合物（NEC）是此过程中的关键参与者。使用 在体外模型系统中，我们先前发现NEC是一种完整的，病毒编码的膜 在核包膜上运行的发芽机。但是，理解核的主要障碍 出口是缺乏了解NEC如何产生膜曲率的知识。这 这项研究的长期目标是阐明疱疹病毒核出口的详细机制，均为 获得对这个不寻常过程的基本知识，并确定和表征新的目标 抗病毒治疗设计。该提议是由中央假设驱动的，基于大量 初步数据，即NEC/膜相互作用和NEC寡聚为外套都是主要的 驱动力，使负膜曲率形成和萌芽。该提议的目的 是通过表征来系统地剖析疱疹样品中的NEC发芽机制（HSV） 必需的蛋白质/蛋白质和蛋白质/膜相互作用以及通过使用A 多学科方法，其中包括冷冻电子显微镜和 电子自旋共振。拟议作品的科学前提是，全面的解剖 NEC介导的负膜曲率的形成对于揭开异常的必要 疱疹病毒出口的机制和制定阻止其策略的机制。除了病毒之外，这项研究将 一般而言，我们对膜变形机制的机理有限理解。这 提案具有创新性，因为它研究了一种不寻常的机制，以原始假设为指导，并且 采用新颖的方法。该提议很重要，因为它旨在提高我们的机械 了解病毒复制周期中的基本步骤，目的是确定新目标 治疗性干预措施，因为它提供了开发负曲率模型的机会 编队，目前是黑匣子。