Defining picornaviral replication complexes by molecular virology and state-of-the art imaging - Novel strategies for disease control

通过分子病毒学和最先进的成像定义小核糖核酸病毒复制复合物 - 疾病控制的新策略

• 批准号: 2271141
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2271141
• 关键词: Defining; picornaviral; replication; complexes; molecular

Background:Picornaviruses are responsible for a number of serious diseases, including polio, foot-and-mouth disease (FMDV) and some emerging diseases, e.g. human hand foot and mouth disease. There is an urgent need to develop new therapeutic strategies to address the continuing issue of picornavirus infection. Genome replication, a key therapeutic target, is usually sequestered within 'replicationcomplexes', thought to improve efficiency and protect this vulnerable part of the lifecycle. However, for picornaviruses, these are poorly defined. Objectives:The project is part of a larger research portfolio designed to improve our understanding of important molecular processes at a fundamental level and to develop new picornavirus therapies, supported by research grants from BBSRC, MRC and WHO/Gates foundation and collaborators (UK and overseas).Novelty:This project will focus on FMDV, many aspects of the replication of this virus are poorly understood. To our knowledge, no similar work is being undertaken elsewhere.Timeliness:We have recently established a reciprocal complementation system to study the cross talk between FMDV genomes [Herod et al, 2015,2016,2017]. These studies have highlighted the importance of multiple co-infection in viral replication. We will exploit this system here, see below.Experimental approach:Our work employs replicons - mini-genomes that allow the study of replication in a safe virus-free way which is highly convenient for the study of the mechanics of genome replication. Using reciprocal complementation (together with state-of the art imaging and structural techniques available in-house) we can dissect the functions of both the RNA and protein components of replication complex. i.e.1. Identifying novel roles of the essential FMDV non-structural proteins and precursors in replication, focusing on the polymerase (3D) and primer of replication (VPg) using mutagenesis and replication inhibitors.2. Investigating the development and structure of sites of replication (replication factories) in a time-resolved manner e.g. by PALM/STORM and EM.

背景：小核糖核酸病毒是引起许多严重疾病的病原，包括脊髓灰质炎、口蹄疫和一些新出现的疾病，如人类手足口病。迫切需要开发新的治疗策略来解决持续存在的小核糖核酸病毒感染问题。基因组复制是一个关键的治疗靶点，通常被隔离在“复制复合体”中，被认为可以提高效率并保护生命周期中的这一脆弱部分。然而，对于小核糖核酸病毒，这些定义很差。目的：该项目是一个更大的研究组合的一部分，旨在提高我们的理解重要的分子过程在一个基本的水平，并开发新的小核糖核酸病毒疗法，由研究赠款的支持，从BBSRC，MRC和WHO/盖茨基金会和合作者（英国和海外）。新奇：该项目将集中在口蹄疫病毒，这种病毒的复制的许多方面知之甚少。据我们所知，其他地方没有进行类似的工作。时间：我们最近建立了一个互惠互补系统来研究FMDV基因组之间的串扰[Herod et al，2015，2016，2017]。这些研究强调了多重共感染在病毒复制中的重要性。实验方法：我们的工作采用了复制子-微型基因组，它允许以安全无病毒的方式研究复制，这对于研究基因组复制的机制非常方便。利用互补作用（结合内部现有的最先进的成像和结构技术），我们可以剖析复制复合物的RNA和蛋白质组分的功能。即1.利用诱变技术和复制引物技术，对口蹄疫病毒复制过程中的聚合酶（3D）和复制引物（VPg）进行了研究，确定了口蹄疫病毒非结构蛋白和前体蛋白在复制过程中的新作用.以时间分辨的方式调查复制站点（复制工厂）的发展和结构，例如通过PALM/STORM和EM。