Impact of host protein dynamics on picornavirus RNA 5’ noncoding region functions

宿主蛋白动态对小核糖核酸病毒 RNA 5â非编码区功能的影响

• 批准号: 9321499
• 负责人: Bert L Semler
• 金额: $ 3.99万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9321499
• 关键词: AUF1A protein; Address; Antiviral Response; Binding; Cell Culture Techniques; Cell Extracts; Cell Line; Cell physiology; Cells; Cleaved cell; Common Cold; Complex; Coxsackie Viruses; Cytoplasm; Defense Mechanisms; Elements; Embryo; Encephalitis; Ensure; Environment; Family; Family Picornaviridae; Fibroblasts; Genetic; Genomics; Health; Hepatitis; High Prevalence; Host Defense Mechanism; Human; Human poliovirus; Hydroxyl Radical; In Vitro; Infection; Instruction; Internal Ribosome Entry Site; Lead; Lung; Measurement; Mediating; Meningitis; Modeling; Mus; Myocarditis; Names; Nature; Play; Poliomyelitis; Process; Protein Dynamics; Proteins; Proteome; Proteomics; RNA; RNA Binding; RNA Stability; RNA Viruses; RNA chemical synthesis; RNA replication; RNA, Ribosomal, 18S; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Regulatory Element; Rhinovirus; Ribosomal RNA; Ribosomes; Role; Site; Testing; Therapeutic; Translation Initiation; Translations; Viral; Viral Physiology; Viral Proteins; Virion; Virus; Virus Diseases; Virus Replication; Work; crosslinking and immunoprecipitation sequencing; genomic RNA; human disease; human morbidity; insight; mRNA Decay; novel; pathogen; prevent; research study; translation factor; viral RNA

 DESCRIPTION (provided by applicant): Picornaviruses are major etiological agents of human diseases, including the common cold, poliomyelitis, encephalitis, meningitis, myocarditis, and hepatitis. They are the most common causes of viral illnesses worldwide and the most common causes of infections of humans in the developed world. Yet they are among the most genetically simple of all mammalian viruses, harboring small, positive-sense genomic RNAs with critical regulatory elements contained within their long 5' noncoding regions (NCRs). This apparent conundrum can, in part, be explained by the way these viruses hijack host cell functions and modify the cytoplasmic environment to favor viral replication. Picornaviruses have evolved complex mechanisms for key steps in their replication cycles, including IRES-mediated translation initiation, the switch in template use from translation to RNA replication, and the evasion of host anti-viral responses that all involve, directly or indirectly, the genomic 5' NCR. Unraveling these mechanisms is the over-arching theme of the experiments proposed in this renewal application. Using poliovirus, human rhinovirus, and coxsackievirus in cell culture models and an array of in vitro experimental approaches, this application will address the fundamental nature of viral processes that are directed by the 5' NCRs of genomic RNAs by proposing to (i) determine how host protein SRp20 interacts with the 40S ribosomal subunit to mediate formation of poliovirus IRES-mediated translation initiation complexes, (ii) define the steps used by human rhinovirus to switch template usage from translation to viral RNA synthesis, and (iii) determine the mechanism of AUF1 negative regulation of viral infections. Results from the proposed studies will reveal important insights into how the 5' NCRs of picornavirus RNAs control a myriad of interactions with host and viral proteins, all in the name of survival of a genetically-challenged family of RNA viruses. Such insights should be broadly applicable to other positive-strand RNA viruses that replicate in the cytoplasm of infected human cells and may help to identify unique virus-host interfaces to be developed as targets for anti-viral therapeutics.

 描述（申请人提供）：小核糖核酸病毒是人类疾病的主要病原体，包括普通感冒、脊髓灰质炎、脑炎、脑膜炎、心肌炎和肝炎。它们是全世界病毒性疾病的最常见原因，也是发达国家人类感染的最常见原因。然而，它们是所有哺乳动物病毒中基因最简单的一种，含有小的、正义基因组 RNA，其长 5' 非编码区 (NCR) 中包含关键调控元件。这一明显的难题可以部分地通过这些病毒劫持宿主细胞功能并改变细胞质环境以有利于病毒复制的方式来解释。小RNA病毒在其复制周期的关键步骤中进化出了复杂的机制，包括IRES介导的翻译起始、模板使用从翻译到RNA复制的转换，以及逃避宿主抗病毒反应，所有这些都直接或间接涉及基因组5'NCR。阐明这些机制是本次更新申请中提出的实验的首要主题。在细胞培养模型中使用脊髓灰质炎病毒、人鼻病毒和柯萨奇病毒以及一系列体外实验方法，该应用将解决由基因组 RNA 5' NCR 指导的病毒过程的基本性质，方法是 (i) 确定宿主蛋白 SRp20 如何与 40S 核糖体亚基相互作用以介导脊髓灰质炎病毒 IRES 介导的翻译的形成 起始复合物，(ii) 定义人鼻病毒将模板使用从翻译转换为病毒 RNA 合成的步骤，以及 (iii) 确定 AUF1 负调节病毒感染的机制。拟议研究的结果将揭示小核糖核酸病毒 RNA 的 5' NCR 如何控制与宿主和病毒蛋白的无数相互作用的重要见解，所有这些都是以 一个受到基因挑战的 RNA 病毒家族的生存。这些见解应该广泛适用于在受感染的人类细胞的细胞质中复制的其他正链RNA病毒，并可能有助于识别独特的病毒-宿主界面，将其开发为抗病毒治疗的靶点。