VIRULENCE DETERMINANTS IN THE COXSACKIEVIRUS B3 GENOME

柯萨奇病毒 B3 基因组的毒力决定因素

• 批准号: 8167492
• 负责人: WILLIAM E TAPPRICH
• 金额: $ 1.03万
• 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: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167492
• 关键词: Antiviral Agents; Chemicals; Computer Retrieval of Information on Scientific Projects Database; Computer-Assisted Image Analysis; Coxsackie Viruses; Disease; Elements; Family Picornaviridae; Funding; Genome; Goals; Grant; Indium; Initiator Codon; Institution; Insulin-Dependent Diabetes Mellitus; Internal Ribosome Entry Site; Maps; Mediating; Methods; Modification; Molecular; Myocarditis; Nucleotides; Pancreatitis; Phenotype; Play; RNA; RNA Folding; RNA-Protein Interaction; Recruitment Activity; Research; Research Personnel; Resources; Ribosomes; Role; Site; Solutions; Source; Structure; Techniques; Testing; Translating; United States National Institutes of Health; Vaccines; Variant; Viral; Viral Proteins; Virulence; Virulent; base; insight; mutant; protein complex

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Coxsackievirus B3 (CVB3) is the leading cause of viral myocarditis, causes pancreatitis and plays a role in type I diabetes. Like all picornaviruses, CVB3 translates its RNA genome using an internal ribosome entry site (IRES), whereby ribosomes are recruited directly to an initiation codon by recognizing a highly structured RNA element in the 5' nontranslated region (5'NTR). The overall goal of this research is to understand the structure and function of the 5'NTR and its associated IRES. The specific objective of this proposal is to explore the RNA elements and the RNA-protein interactions that determine virulence in CVB3. The two specific aims of the previous grant were accomplished, confirming the hypothesis that the IRES folds into a stable structure that is required for proper function. As proposed in specific aim 1, the structure of a wild type CVB3 IRES was determined. As proposed in specific aim 2, differences that mediate the virulence phenotype have been mapped. The goal of the current research is to define the minimal RNA elements that determine virulence and explore the RNA-protein interactions that underlie IRES function. Two specific aims will test the hypothesis that small, independently folding RNA structures in the 5'NTR provide recognition features for cellular and viral proteins that together determine viral virulence. In specific aim 1, the solution structure of RNA domains previously identified to be virulence determinants will be determined in virulent and non-virulent variants of the genome. These variants will include naturally occurring sequences, site-directed mutants and chimeric constructs. Our established methods of base-specific chemical modifying agents will probe the accessibility of nucleotides in the folded RNA domains. In specific aim 2 the molecular mechanism of IRES function will be explored by studies of RNA-protein interactions. Probing studies of RNA-protein complexes using our established chemical modification techniques will provide insight into the conformational dynamics and functional states of the 5'NTR. These results will aid in the search for effective antivirals and vaccines, not only for CVB3 but also for a host of other disease-causing picornaviruses.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 Coxsackievivirus B3（CVB3）是病毒心肌炎的主要原因，引起胰腺炎，并在I型糖尿病中发挥作用。 与所有PICORNAVIRES一样，CVB3使用内部核糖体输入位点（IRES）转换其RNA基因组，从而通过在5'非翻译区（5'NTR）中识别出高度结构化的RNA元素，从而将核糖体直接募集到起始密码子中。 这项研究的总体目标是了解5'NTR及其相关的IRES的结构和功能。 该提案的具体目的是探索确定CVB3毒力的RNA元素和RNA蛋白相互作用。 实现了先前赠款的两个具体目标，证实了以下假设：IRES折叠成适当功能所需的稳定结构。 正如特定目标1中提出的那样，确定了野生型CVB3 IRES的结构。 正如特定目标2中提出的那样，介导毒力表型的差异已被映射。 当前研究的目的是定义确定毒力的最小RNA元素，并探索IRES起作用的RNA蛋白质相互作用。 两个具体的目的将检验以下假设：5'NTR中的小，独立折叠的RNA结构为细胞和病毒蛋白提供了识别特征，这些特征共同决定了病毒毒力。 在特定的目标1中，将在基因组的毒和非毒气变体中确定先前确定为毒力决定因素的RNA结构域的溶液结构。 这些变体将包括天然发生的序列，位置为导向的突变体和嵌合构建体。 我们确定的基本化学修饰剂的方法将探测折叠RNA结构域中核苷酸的可及性。 在特定目标2中，IRES功能的分子机制将通过RNA-蛋白质相互作用的研究来探索。 使用我们已建立的化学修饰技术对RNA-蛋白质复合物进行探测研究将提供对5'NTR的构象动力学和功能状态的见解。 这些结果将有助于寻找有效的抗病毒药和疫苗，不仅对于CVB3，而且对于其他许多引起疾病的Picornavires。