Defining the nuclear vs. cytoplasmic proteome during human rhinovirus infections

定义人鼻病毒感染期间的核与细胞质蛋白质组

• 批准号: 9293982
• 负责人: Bert L Semler
• 金额: $ 18.36万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-10 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9293982
• 关键词: Address; Antiviral Agents; Architecture; B-Lymphocytes; Biological Assay; Cell Nucleus; Cell physiology; Cells; Cleaved cell; Code; Confocal Microscopy; Coxsackie Viruses; Cytoplasm; Enterovirus 71; Environment; Event; Family Picornaviridae; Fibroblasts; Growth; Hela Cells; High Prevalence; Human; Human poliovirus; Infection; Internal Ribosome Entry Site; Lung; Mass Spectrum Analysis; Mediating; Modification; Morbidity - disease rate; Nuclear; Nuclear Proteins; Phase; Play; Prevalence; Protein Biosynthesis; Proteins; Proteome; Proteomics; RNA Interference; RNA Viruses; RNA chemical synthesis; RNA replication; RNA-Binding Proteins; Rhinovirus; Role; Scheme; Testing; Therapeutic; Time; Translating; Translations; Vesicle; Viral; Virion; Virus; Virus Diseases; Virus Replication; Western Blotting; experimental study; genomic RNA; human morbidity; insight; novel; pathogen; trafficking; viral RNA; virus host interaction

Picornaviruses are major human pathogens, and their worldwide prevalence is responsible for some of the highest morbidities of all known viruses. Due to the limited coding capacity of picornavirus genomic RNAs, host proteins play critical roles during viral translation and RNA replication in the cytoplasm of infected cells. A number of these proteins are normally localized to the nucleus of uninfected human cells. The experiments proposed in this application aim to define the changing protein landscape in human rhinovirus infected cells that is brought about by viral-induced modifications of the nucleo-cytoplasmic trafficking machinery of the host cell. The proposal will explore the unique mechanisms that rhinovirus has evolved to take advantage of host nuclear proteins which play key roles in IRES-mediated translation and viral RNA synthesis during its intracellular replication cycle. By interacting with normally nuclear-resident proteins during its cytoplasmic replication cycle, rhinovirus expands the repertoire of host cell functions it can hijack for its own purposes. Using an unbiased proteomics approach, this application will address the hypothesis that human rhinovirus induces a re-partitioning of host cell proteins between the nucleus and the cytoplasm to potentiate steps in its replication cycle. The proposed experiments will expand upon a recent discovery that rhinovirus infection of human lung fibroblast cells results in a different set of virus-host interactions compared to what is observed during a rhinovirus or poliovirus infection of HeLa cells. Two experimental aims will be carried out to discover if human rhinovirus uses a distinct set of host proteins and mechanisms during replication in lung cells aided, in part, by the novel viral-induced redistribution of resident nuclear proteins to the cytoplasm of infected cells: (1) Generate a nuclear versus cytoplasmic human cell proteome during a human rhinovirus infection, and (2) Identify candidate host proteins and determine their functional significance for human rhinovirus replication. Overall, determining how rhinoviruses and other picornaviruses take advantage of altered nucleo-cytoplasmic trafficking should reveal new mechanistic insights into RNA virus replication and identify novel targets for antiviral therapeutics at the virus-host interface.

小核糖核酸病毒是人类的主要病原体，其在世界范围内的流行是导致人类免疫缺陷的主要原因。 是所有已知病毒中发病率最高的由于有限的编码能力， 小核糖核酸病毒基因组RNA，宿主蛋白在病毒翻译和RNA合成过程中起关键作用。 在受感染细胞的细胞质中复制。这些蛋白质中的许多通常定位于 转移到未受感染的人体细胞核上本申请中提出的实验旨在 定义人类鼻病毒感染细胞中蛋白质的变化， 通过病毒诱导的宿主细胞核质运输机制的修饰。 该提案将探索鼻病毒进化的独特机制， 在IRES介导的翻译和病毒RNA中起关键作用的宿主核蛋白 在其细胞内复制周期期间合成。通过与正常核居民的互动， 在其细胞质复制周期中，鼻病毒扩大了宿主细胞 它可以为自己的目的劫持的功能。使用无偏见的蛋白质组学方法， 应用将解决人鼻病毒诱导宿主重新分配的假设 细胞核和细胞质之间的蛋白质，以加强其复制周期中的步骤。 拟议的实验将扩大最近的发现，鼻病毒感染的 人肺成纤维细胞导致一组不同的病毒-宿主相互作用， 在鼻病毒或脊髓灰质炎病毒感染HeLa细胞期间观察到。两个实验目标将 以发现人类鼻病毒是否使用一组不同的宿主蛋白质， 在肺细胞中复制过程中的机制，部分是由新的病毒诱导的 （1）在感染细胞的细胞质中重新分布常驻核蛋白： 人鼻病毒感染期间的核与细胞质人细胞蛋白质组，和（2） 鉴定候选宿主蛋白并确定其对人鼻病毒的功能意义 复制的总的来说，确定鼻病毒和其他小核糖核酸病毒如何利用 改变的核质运输应该揭示RNA病毒的新机制 复制，并在病毒-宿主界面上鉴定抗病毒治疗的新靶点。