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Functional development of picornavirus-induced membranes throughout infection

小核糖核酸病毒诱导的细胞膜在感染过程中的功能发育

基本信息

批准号：
10681226
负责人：
William T Jackson
金额：
$ 38.63万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10681226
关键词：
Amino Acids Autophagocytosis Autophagosome Biochemical Capsid Capsid Proteins Cell membrane Cell physiology Cells Cellular Membrane Comparative Study Complex Coxsackie Viruses Detection Development Disease Disease Outbreaks Endosomes Enterovirus Enterovirus 68 Environment Experimental Designs Family Picornaviridae Foot-and-Mouth Disease Generations Human poliovirus Immune Evasion Infection Internet Knowledge Life Cycle Stages Lipids Membrane Membrane Proteins Methods Modeling Modification Mouth Diseases Myocarditis Neurodegenerative Disorders Pathway interactions Poliovirus Vaccines Production Proteins Public Health RNA replication Regulation Research Respiratory distress Series Signal Transduction Site Starvation Stress Testing Therapeutic Tube Vesicle Viral Viral Nonstructural Proteins Viral Proteins Virion Virus Virus Replication acute flaccid myelitis experimental study human disease inhibition of autophagy inhibitor insight member new outbreak pathogen prevent protein aggregation therapeutic development therapeutic target viral RNA

项目摘要

Enteroviruses are among the most severe causes of human disease, and recent outbreaks of emerging enteroviruses such as types D68 and 71 have highlighted the need to understand more about these pathogens. Our comparative studies of poliovirus (PV), coxsackievirus B3, and now EV-D68 have shown many commonalities in the life-cycles of these viruses. We have focused our studies on how members of the Enterovirus genus trigger a cellular pathway known as autophagy to promote production of infectious virus. Autophagy is a constitutive degradative cellular process required for turnover of damaged vesicles, aggregated proteins, and other spent cellular components. During times of stress, including amino acid starvation, organismal development, and infection, autophagy is up-regulated. Autophagic vesicles, thought to be derived from the complex web of viral RNA replication membranes, can be observed during mid-to-late infection. We and others have demonstrated that viral proteins specifically induce autophagic signaling and autophagosome formation. These autophagosomes acidify, promoting maturation of the viral capsid and release of newly formed infectious viruses, often encased in these cell-derived membranes, from the cell. In this proposal, we outline a comprehensive plan to understand generation of membranes during infection, from the initial RNA replication membranes to the autophagosomes promoting maturation of virions to the single-membraned virus- containing vesicles being released from the cell. Understanding this entire pathway, the latter parts of which have only been identified within the past few years, will provide an understanding of how these viruses replicate, mature, release from cells, and evade immune detection. In Specific Aim I, we will analyze how specific viral non-structural proteins initiate autophagosome formation at the early stages of infection and regulate downstream steps in autophagosome formation, acidic maturation, and virus release. In Specific Aim II, we will investigate the viral and host requirements for development of viral RNA replication membranes, and the mechanism of formation of autophagosomes from these convoluted membrane structures, during infection. These experiments can be carried out in the absence of host proteins from the autophagic pathway and other cellular pathways. In addition, we have shown that acidic autophagosomes promote maturation of the virion by inducing cleavage of the capsid protein VP0 into VP2 and VP4. We propose a biochemical approach to identifying the factor(s) contained within autophagosomes required for infectious maturation of viral capsids. There is a major research gap in understanding development of enterovirus-induced membranes, from the initial signals sent by virus proteins at the beginning of infection, to the release of virus-containing vesicles. This proposal will address that need by investigating each step in this pathway from beginning - initiation of RNA replication membranes - to end - the release of virus-containing vesicles.

肠道病毒是人类疾病最严重的原因之一，最近爆发的新出现的肠道病毒，如D68和71型，突显了更多了解这些病原体的必要性。我们对脊髓灰质炎病毒(PV)、柯萨奇病毒B3和现在的EV-D68的比较研究表明，这些病毒在生命周期中有许多共同之处。我们的研究重点是肠道病毒属成员如何触发一种称为自噬的细胞途径，以促进传染性病毒的产生。自噬是一种结构性降解的细胞过程，需要周转受损的囊泡、聚集的蛋白质和其他废弃的细胞成分。在压力时期，包括氨基酸饥饿、生物体发育和感染，自噬被上调。自噬小泡被认为来自病毒RNA复制膜的复杂网络，在中晚期感染期间可以观察到。我们和其他人已经证明，病毒蛋白特异性地诱导自噬信号和自噬小体的形成。这些自噬小体酸化，促进病毒衣壳的成熟和新形成的传染性病毒的释放，这些病毒通常被包裹在这些细胞衍生的膜中，从细胞释放出来。在这项提案中，我们概述了一个全面的计划，以了解感染过程中膜的生成，从最初的RNA复制膜到促进病毒粒子成熟的自噬小体，再到从细胞中释放的含有病毒的单膜囊泡。了解这一整个途径--其后期部分在过去几年才被发现--将提供对这些病毒如何复制、成熟、从细胞中释放以及逃避免疫检测的理解。在特定的目标I中，我们将分析特定的病毒非结构蛋白如何在感染的早期阶段启动自噬小体的形成，并调节自噬小体形成、酸性成熟和病毒释放的下游步骤。在特定的目标II中，我们将研究病毒和宿主对病毒RNA复制膜发育的要求，以及在感染过程中从这些卷曲的膜结构形成自噬小体的机制。这些实验可以在没有来自自噬途径和其他细胞途径的宿主蛋白的情况下进行。此外，我们还表明，酸性自噬小体通过诱导衣壳蛋白VP0裂解成VP2和VP4来促进病毒粒子的成熟。我们提出了一种生化方法来鉴定自噬体内包含的因子(S)，这是病毒衣壳感染性成熟所必需的。在了解肠道病毒诱导的膜的发育方面，从感染开始时病毒蛋白发出的初始信号，到含有病毒的囊泡的释放，存在着重大的研究空白。这项建议将通过研究这一途径中从开始-启动RNA复制膜-到结束-释放含病毒小泡的每一步来满足这一需求。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Autophagy-Associated Proteins Control Ebola Virus Internalization Into Host Cells.

DOI：
10.1093/infdis/jiy294
发表时间：
2018-11-22
期刊：
The Journal of infectious diseases
影响因子：
0
作者：
Shtanko O;Reyes AN;Jackson WT;Davey RA
通讯作者：
Davey RA

SIRT-1 is required for release of enveloped enteroviruses.

DOI：
10.7554/elife.87993
发表时间：
2023-10-18
期刊：
eLife
影响因子：
7.7
作者：
Jassey A;Logue J;Weston S;Wagner MA;Galitska G;Miller K;Frieman M;Jackson WT
通讯作者：
Jackson WT

Poliovirus-induced changes in cellular membranes throughout infection.

DOI：
10.1016/j.coviro.2014.09.007
发表时间：
2014-12
期刊：
CURRENT OPINION IN VIROLOGY
影响因子：
5.9
作者：
Jackson, William T.
通讯作者：
Jackson, William T.

Starvation after infection restricts enterovirus D68 replication.

DOI：
10.1080/15548627.2022.2062888
发表时间：
2023-01
期刊：
AUTOPHAGY
影响因子：
13.3
作者：
Jassey, Alagie;Wagner, Michael A.;Galitska, Ganna;Paudel, Bimal;Miller, Katelyn;Jackson, William T.
通讯作者：
Jackson, William T.

Enterovirus D68 capsid formation and stability requires acidic compartments.

肠道病毒 D68 衣壳的形成和稳定性需要酸性隔室。