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: 7885216
• 负责人: Nora M. Chapman
• 金额: $ 29.22万
• 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 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7885216
• 关键词: Acute; Adult; Affect; Binding; Binding Sites; Biology; Cell Culture Techniques; Cell Nucleus; Cells; Child; Chronic; Chronic Disease; Complement component C1s; Complex; Coxsackie B Viruses; Cytoplasm; Dilated Cardiomyopathy; Disease; Enterovirus; Enterovirus Infections; Environment; Etiology; Event; Face; Family Picornaviridae; Generations; Genome; Genomics; Goals; Growth; Heart; Higher Order Chromatin Structure; Human; Human poliovirus; Immune; Immune response; Infection; Knowledge; Laboratories; Lead; Life Style; Link; Long-Term Effects; Lytic; Mus; Mutate; Myocarditis; Neurologic; 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; 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RNA病毒在适应新环境方面非常成功。我们将在这个项目中探索这样一种新发现的适应性，确定对复制效率至关重要的基因组区域的丢失如何使肠道病毒适应不同的细胞内环境。肠道病毒被认为具有快速裂解的特点，可引起短期感染。然而，B 组柯萨奇病毒 (CVB) 可以删除基因组 5' 末端 (TD) 的序列，但仍保持功能，最重要的是，在感染后在心脏或细胞培养物中长期存在，这一发现为挑战这一传统观点提供了依据。鉴于肠道病毒 RNA 的 5' 末端参与病毒复制的多种功能，从基因组 5' 末端删除多达 49 个核苷酸对于病毒来说应该是一个可怕的事件，事实确实如此。除其他外，病毒复制显着减慢，正链 RNA 合成受​​损导致受感染细胞中的阳性病毒 RNA 与阴性病毒 RNA 的水平接近 1 比 1。该提案的目的是描述 hnRNPC1 在 CVB-TD 群体建立机制中的作用。我们假设，在宿主细胞定义的情况下（静止细胞中缺乏细胞质 hnRNPC1），会出现另一种形式的正链病毒 RNA 合成起始，导致正链基因组的 5' 端缺失。提出了一个单一的具体目标来实现这一目标：我们将确定静止细胞中是否由于病毒转录起始的改变而发生一定范围的末端缺失，该范围是否受到5'非翻译区结构域I部分的要求的限制，并且我们将检查CVB-TD复制复合物的结构和蛋白质。如果 hnRNPC1 不能结合 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，以及肠道病毒蛋白引物的野生型尿苷酰化过程在多大程度上参与 CVB3-TD 正链 RNA 合成的起始。总而言之，这项工作将显着扩展我们对这种令人着迷的肠道病毒生活方式的了解，提供对肠道病毒持续存在并导致人类慢性疾病的新机制的见解。 公共卫生相关性：本研究旨在确定持续性肠道病毒感染的产生条件和过程。这些心脏感染与心脏功能的长期影响有关，包括扩张性心肌病和慢性心肌炎等衰弱性疾病。了解这些变异病毒如何发生选择以及这些病毒如何复制将为开发治疗方法和检测这些病毒在心脏、神经和胰腺疾病中的基础提供基础。