Equine rhinitis A virus as a model for foot-and-mouth-disease virus: mechanism of RNA release and membrane penetration

马鼻炎A病毒作为口蹄疫病毒模型：RNA释放和膜穿透机制

• 批准号: BB/E00931X/1
• 负责人: David Rowlands
• 金额: $ 40.53万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE00931X%2F1
• 关键词: Equine; rhinitis; virus; model; foot

Summary Foot-and-mouth disease virus (FMDV) is one of the most important pathogens affecting agricultural livestock. It is endemic in much of the world, especially in the developing world where its presence restricts the ability to export animal products to disease free countries and so adversely affects rural economies. The virus is one of the most infectious agents known and its ability to cause economic and social havoc was amply demonstrated by the outbreak in the UK in 2001 which cost the country ~£10 billion. Because of the danger of spreading infection only a small number of laboratories world/wide have the containment facilities required to be licensed for work with the disease. This clearly restricts the amount of research that can be conducted, especially on fundamental aspects of the virus and its replication. Recently, equine rhinitis A virus (ERAV) was classified into the same genus, the aphthoviridae, as FMDV on the basis of its genetic similarity. ERAV causes relatively mild symptoms in horses and is not subject to the severe restrictions applied to FMDV and it can be handled in general virology laboratories. In this application we wish to investigate the initial steps of the virus infection process using ERAV in the anticipation that it will be a useful model for FMDV itself. Both ERAV and FMDV are members of the picornavirus family and one of the least well understood steps in the infection process by any of these viruses is how the virus particle is uncoated and transports its RNA genome across the cell membrane to initiate an infection. We have been studying this process, in collaboration with other laboratories, using poliovirus (PV), which is the best understood of all picornaviruses. From this work, a model of how the infection process is achieved by PV is slowly evolving. It appears to be an elegant and sophisticated mechanism involving a series of intermediate particles. However, the known properties of aphthovirus particles make it difficult to extrapolate the PV model to these viruses. We intend to apply the novel techniques that we have developed in recent years for studying PV to investigate in detail the mechanism of cell entry by ERAV. These methods use artificial lipid membrane structures to mimic cellular membranes in such a way that they can be finely controlled and examined in the laboratory. We will attempt to mimic the infectious process using these systems and examine the intermediate structures formed during the uncoating of the virus and their effects on the properties of the membranes themselves. In this way we hope to gain insight into how the virus uncoats and projects its RNA across an artificial equivalent of a cell membrane. In addition, we will determine the crystal structures of the native virus and potential cell-entry intermediate particles by X-ray diffraction in collaboration with colleagues in Oxford. As we gain a better understanding of the infection process with ERAV we plan to extend the work to include FMDV via our collaborators at the Institute for Animal Health, Pirbright. Although these studies are of immediate academic interest, a better understanding of the infection process may also have more practical value in the longer term. The precise requirements and mechanisms used by viruses are important in determining tissues and species specificity of infection and so are important for understanding host range of the virus. In addition, better understanding of the infectious process can lead to the development of pharmaceutical approaches to interfere with it, as has been shown for HIV.

口蹄疫病毒（Foot-and-mouth disease virus，FMDV）是影响农业家畜的重要病原之一。它在世界许多地方流行，特别是在发展中国家，它的存在限制了向无病国家出口动物产品的能力，从而对农村经济产生不利影响。该病毒是已知的最具传染性的病原体之一，其造成经济和社会破坏的能力在2001年英国爆发的疫情中得到了充分证明，该疫情使该国损失了约100亿英镑。由于有传播感染的危险，全世界只有少数实验室拥有必须获得许可才能从事这种疾病工作的隔离设施。这显然限制了可以进行的研究的数量，特别是对病毒及其复制的基本方面的研究。最近，马鼻炎A病毒（ERAV）被归类为同一属，口蹄疫病毒，口蹄疫病毒的遗传相似性的基础上。ERAV在马中引起相对轻微的症状，并且不受适用于FMDV的严格限制，并且可以在普通病毒学实验室中处理。在本申请中，我们希望调查的病毒感染过程中使用ERAV的预期，它将是一个有用的模型FMDV本身的初始步骤。ERAV和FMDV都是小核糖核酸病毒家族的成员，这些病毒感染过程中最不了解的步骤之一是病毒颗粒如何未包衣并将其RNA基因组转运穿过细胞膜以引发感染。我们一直在与其他实验室合作，使用脊髓灰质炎病毒（PV）研究这一过程，这是所有小核糖核酸病毒中最好的理解。从这项工作中，一个如何通过PV实现感染过程的模型正在慢慢发展。这似乎是一个优雅而复杂的机制，涉及一系列中间粒子。然而，口疮病毒颗粒的已知特性使得难以将PV模型外推到这些病毒。我们打算应用新的技术，我们已经开发了近年来研究PV详细调查ERAV进入细胞的机制。这些方法使用人工脂质膜结构来模拟细胞膜，以便在实验室中对其进行精细控制和检查。我们将尝试使用这些系统来模拟感染过程，并检查在病毒脱壳过程中形成的中间结构及其对膜本身性质的影响。通过这种方式，我们希望能够深入了解病毒是如何将其RNA剥离并投射到人工细胞膜上的。此外，我们将与牛津大学的同事合作，通过X射线衍射确定天然病毒和潜在细胞进入中间颗粒的晶体结构。随着我们对ERAV感染过程的更好理解，我们计划通过我们在Pirbright动物卫生研究所的合作者将工作扩展到包括FMDV。虽然这些研究具有直接的学术意义，但从长远来看，更好地了解感染过程也可能具有更大的实用价值。病毒的确切需求和机制对于确定感染的组织和种属特异性非常重要，因此对于了解病毒的宿主范围也很重要。此外，更好地了解感染过程可以导致开发药物方法来干扰它，如艾滋病毒所示。

项目成果

Picornaviruses.

• DOI: 10.1007/82_2010_37
• 发表时间: 2010
• 期刊: CURRENT TOPICS IN MICROBIOLOGY AND IMMUNOLOGY
• 作者: Tuthill, Tobias J.;Groppelli, Elisabetta;Hogle, James M.;Rowlands, David J.
• 通讯作者: Rowlands, David J.

Picornavirus RNA is protected from cleavage by ribonuclease during virion uncoating and transfer across cellular and model membranes.

• DOI: 10.1371/journal.ppat.1006197
• 发表时间: 2017-02
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Groppelli E;Levy HC;Sun E;Strauss M;Nicol C;Gold S;Zhuang X;Tuthill TJ;Hogle JM;Rowlands DJ
• 通讯作者: Rowlands DJ

Limits of structural plasticity in a picornavirus capsid revealed by a massively expanded equine rhinitis A virus particle.

• DOI: 10.1128/jvi.01979-13
• 发表时间: 2014-06
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Bakker SE;Groppelli E;Pearson AR;Stockley PG;Rowlands DJ;Ranson NA
• 通讯作者: Ranson NA