VIRULENCE DETERMINANTS IN PICORNAVIRUS GENOMIC RNA

小核病毒基因组 RNA 的毒力决定因素

• 批准号: 7627608
• 负责人: WILLIAM E TAPPRICH
• 金额: $ 4.82万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7627608
• 关键词: 5' Untranslated Regions; Antiviral Agents; Attenuated; Chemicals; Computer Retrieval of Information on Scientific Projects Database; Coxsackie Viruses; Disease; Family Picornaviridae; Funding; Genome; Genomics; Goals; Grant; Indium; Initiator Codon; Institution; Insulin-Dependent Diabetes Mellitus; Internal Ribosome Entry Site; Maps; Mediating; Methods; Modeling; Myocarditis; Nucleotides; Pancreatitis; Phenotype; Phylogenetic Analysis; Play; RNA; Recruitment Activity; Research; Research Personnel; Resources; Ribosomes; Role; Solutions; Source; Structure; Testing; Translating; United States National Institutes of Health; Vaccines; Viral; Virulence; Virulent; Virus; Virus Diseases; base; dimethyl sulfate; insight; kethoxal; mutant

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 clearly plays a role in type I diabetes. Like other 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'UTR) of the genome. The overall goal of this research is to understand the structure and function of the 5'UTR and its associated IRES. The specific objective of this proposal is to explore the 5'UTR and IRES found in coxsackievirus B3 (CVB3). Structural integrity of the IRES is critically important for viral multiplication and also for virulence. Given the central role of the IRES to viral infection it is essential to understand the details of IRES structure and function. For the picornavirus IRES, a secondary structure model has been proposed based upon phylogenetic comparison, but this model has not been tested experimentally. The goal of this research is to determine the solution structure of the CVB3 IRES from both virulent wild type and attenuated mutant viruses. Two specific aims will test the hypothesis that the IRES folds into a stable structure that is required for proper function. In specific aim 1 the structure of a wild type CVB3 IRES will be determined. Base-specific chemical modifying agents such as dimethyl sulfate and kethoxal will be used to probe the accessibility of nucleotides in the folded IRES RNA. Results of the chemical probing will experimentally determine the structure of the IRES. In specific aim 2 structural differences that mediate the virulence phenotype in CVB3 will be mapped. Applying chemical probing methods to attenuated mutant strains of the virus will reveal the critical structural changes that underlie virulence. Understanding the structure, and structural dynamics of the CVB3 IRES will provide insight into targets for inactivating this virus. 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，并且在I型糖尿病中显然起作用。 与其他PICORNAVIRASE一样，CVB3使用内部核糖体入口位点（IRES）转换其RNA基因组，从而通过识别基因组的5'非翻译区（5'UTR）中的高度结构化的RNA元件，直接募集核糖体。 这项研究的总体目标是了解5'UTR及其相关的IRES的结构和功能。 该提案的具体目标是探索Coxsackievivirus B3（CVB3）中发现的5'UTR和IRES。 IRES的结构完整性对于病毒乘法和毒力至关重要。 考虑到IRES对病毒感染的核心作用，必须了解IRES结构和功能的细节。 对于Picornavirus IRES，已经基于系统发育比较提出了二级结构模型，但是该模型尚未通过实验进行测试。 这项研究的目的是确定来自有毒野生型和减弱突变病毒的CVB3 IRES的溶液结构。 两个具体的目的将检验以下假设：IRES折叠成适当功能所需的稳定结构。 在特定目标中，将确定野生型CVB3 IRES的结构。 碱基特异性化学修饰剂（例如硫酸二甲基二甲基二甲基和酮）将用于探测折叠IRES RNA中核苷酸的可及性。 化学探测的结果将通过实验确定IRES的结构。 在特定的目标中，将映射介导CVB3中毒力表型的结构差异。 将化学探测方法应用于病毒的突变菌株中，将揭示毒力构成的关键结构变化。 了解CVB3 IRES的结构和结构动力学将为灭活该病毒的靶标提供洞察力。 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，而且对于许多其他引起picornavires的疾病。