5' End Genomic Deletions and Enterovirus Persistence

5 端基因组缺失和肠道病毒持久性

• 批准号: 8206597
• 负责人: Nora M. Chapman
• 金额: $ 29.4万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-15 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206597
• 关键词: Acute; Adult; Affect; Binding; Binding Sites; Biology; Cell Culture Techniques; Cell Nucleus; Cells; Child; Chronic; Chronic Disease; Complex; Coxsackie B Viruses; Cytoplasm; Dilated Cardiomyopathy; Disease; Enterovirus; Enterovirus Infections; Environment; Etiology; Event; Family Picornaviridae; Generations; Genome; Genomics; Goals; Growth; Heart; Heart Diseases; Higher Order Chromatin Structure; Human; Human poliovirus; Immune; Immune response; Infection; Knowledge; Laboratories; Lead; Life Style; Link; Long-Term Effects; Lytic; Mus; Mutate; Myocarditis; Nuclear; Nucleotides; Pancreatic Diseases; Polioviruses; Population; Primary Cell Cultures; Process; Production; Proteins; RNA; RNA Viruses; RNA chemical synthesis; RNA replication; Reporting; Role; Serotyping; Site; Time; Tissues; Variant; Viral; Viral Physiology; Virion; Virus; Work; base; fascinate; heart function; human disease; insight; nervous system disorder; novel; protein structure; public health relevance; sound; viral RNA; viral detection

DESCRIPTION (provided by applicant): The mutable RNA viruses are marvelously successful at adapting to exploit new environments. We will explore just such a newly discovered adaptation in this project, determining how the loss of a genomic region that is vital to replicational efficiency, allows an enterovirus to adapt to a different intracellular environment. Enteroviruses have been viewed as characteristically rapidly lytic, causing short-term infections. However, the discovery that group B coxsackieviruses (CVB) can delete sequence from the genomic 5' terminus (TD), yet maintain function and most importantly, persist for long periods of time post-infection in the heart or cell culture, has provided the grist with which to challenge that traditional view. Given that the 5' terminus of enterovirus RNA is involved in numerous functions for viral replication, deletion of as many as 49 nucleotides from the 5' end of the genome should be a dire event for the virus and indeed, it is. Among other things, viral replication slows dramatically and crippled positive strand RNA synthesis results in near 1 to 1 levels of positive to negative viral RNA in infected cells. The objective of this proposal is to characterize the role of hnRNPC1 in the mechanism by which CVB-TD populations become established. We hypothesize that under circumstances defined by the host cell - a lack of cytoplasmic hnRNPC1 in quiescent cells - an alternative form of positive strand viral RNA synthesis initiation occurs that leads to 5' end deletions of the positive strand genome. A single specific aim is proposed to accomplish this objective: we will determine whether a range of a terminal deletions occurs in quiescent cells due to altered viral transcriptional initiation, whether that range is limited by a requirement for part of structural domain I of the 5'nontranslated region and we will examine the structure and proteins of the CVB-TD replication complex. We will determine how selective lowering of cytoplasmic hnRNPC1 affects wildtype and TD CVB replication, if hnRNPC1 fails to bind the mutated 3' terminus of CVB- TD negative strand RNA and to what extent the wildtype process of uridylylation of the enterovirus protein primer is involved in CVB3-TD initiation of positive strand RNA synthesis. Cumulatively, this work will significantly expand our knowledge of this fascinating enterovirus lifestyle, providing insight into a novel mechanism by which enteroviruses can persist and cause chronic disease in humans. PUBLIC HEALTH RELEVANCE: This study is to determine the conditions and processes for generation of persistent enterovirus infections. These infections in hearts are associated with long term effects upon heart function including debilitating conditions such as dilated cardiomyopathy and chronic myocarditis. Understanding how selection of these variant viruses occurs and how these viruses replicate will provide the basis for developing treatments and detection of these viruses in heart, neurological and pancreatic disease.

描述（由申请人提供）：可变的RNA病毒在适应新环境方面非常成功。我们将在该项目中探索这种新发现的适应性，以确定对复制效率至关重要的基因组区域的损失，使肠病毒能够适应不同的细胞内环境。肠病毒已被视为特征迅速的裂解，导致短期感染。然而，发现B组Coxsackievievievieviruses（CVB）可以从基因组5'末端（TD）中删除序列，但仍保持功能，最重要的是，在心脏或细胞培养后长时间持续存在持续存在的序列，这为卫生提供了纪念，这为传统观点提供了挑战。鉴于5'肠病毒RNA的5'末端参与了许多用于病毒复制的功能，因此从基因组的5'末端删除了多达49个核苷酸，应该是病毒的可怕事件，实际上是。除其他外，病毒复制会大大减慢和残废的阳性链RNA合成导致感染细胞中接近1至1个阳性至阴性病毒RNA的水平。该建议的目的是表征HNRNPC1在建立CVB-TD种群的机制中的作用。我们假设在宿主细胞定义的情况下 - 静态细胞中缺乏细胞质HnRNPC1 - 发生阳性链病毒RNA合成起始的另一种形式，导致阳性链基因组的5'端缺失。提出了一个特定的目的来实现这一目标：我们将确定由于病毒转录启动的改变而导致的静态细胞中是否发生了一系列末端缺失，该范围是否受到5'nontranslatslated区域I的部分结构域I的要求限制，并且我们将检查CVB-TD复制复合复合复合复合复合物的结构和蛋白质。如果HNRNPC1未能结合CVB-TD负链RNA的突变3'末端，我们将确定细胞质HNRNPC1的选择性降低如何影响野生型和TD CVB复制。累积地，这项工作将大大扩展我们对这种迷人的肠病毒生活方式的了解，从而洞悉一种新型机制，肠病毒可以持续存在并引起人类慢性疾病。 公共卫生相关性：这项研究是为了确定产生持续性肠病毒感染的条件和过程。心脏中的这些感染与对心脏功能的长期影响有关，包括使心肌病和慢性心肌炎等衰弱状况。了解这些变体病毒的选择是如何发生的，以及这些病毒如何复制为开发疗法和在心脏，神经和胰腺疾病中检测这些病毒的基础。