Structure and dynamics of RNA elements regulating viral aberrant RNA synthesis

调节病毒异常RNA合成的RNA元件的结构和动力学

• 批准号: 10472311
• 负责人: Arend Jan te Velthuis
• 金额: $ 141.23万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-08 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10472311
• 关键词: Amino Acids; Avian Influenza A Virus; Basic Amino Acids; Biochemical; Biophysics; Birds; DNA-Directed RNA Polymerase; Disease Marker; Disease Outbreaks; Domestic Fowls; Elements; Genes; Health; Hemagglutinin; Human; Immune response; Industry; Influenza; Influenza A virus; Innate Immune Response; Knowledge; Length; Membrane Proteins; Methods; Molecular; Nucleotides; Organ; Pathogenicity; RNA; RNA chemical synthesis; Research; Site; Structure; Stuttering; Up-Regulation; Viral; Viral Genome; Viral Hemagglutinins; Virus; Virus Diseases; Virus Replication; influenzavirus; mortality; novel; pandemic influenza; pathogenic virus; prevent; seasonal influenza; viral RNA

SUMMARY Seasonal influenza viruses cause >600 million cases annually and hundreds of billions of dollars in losses. Pandemic and avian influenza viruses present an even greater threat because they can dysregulate our immune response and/or spread to and shut down multiple organs. Current evidence suggests that aberrant viral replication contributes to a dysregulating of the innate immune response and the emergence of highly pathogenic strains from low pathogenic precursors. The transition from a low to highly pathogenic virus involves insertion of multiple basic amino acids in the cleavage site of the viral hemagglutinin (HA) surface protein. In birds, this change in HA allows the virus to spread systemically, resulting in mortality rates of up to 100% in poultry. The molecular mechanism underlying insertion of amino acids in the cleavage site is not well understood, but it may involve stuttering of the RNA polymerase in the HA gene, resulting in nucleotide insertions. The viral RNA polymerase can also delete nucleotides from the viral genome, resulting in shorter aberrant RNAs. Recent studies have shown that pandemic and avian influenza A virus infections produce RNA molecules of about 56-125 nucleotides in length, called mini viral RNAs, and that their synthesis is correlated with the upregulation of disease markers. How the RNA polymerase makes such large deletions in the viral genome is not known. This project will use a novel method to stall the viral RNA polymerase during key steps of viral replication and aberrant RNA synthesis, and use state-of-the-art biochemical, biophysical, and structural approaches to reveal the steps of viral replication and genome encapsidation, as well as aberrant RNA synthesis. In doing so, this project will contribute to a complete mechanistic understanding of influenza replication and answer long-standing, fundamental questions about the emergence of highly pathogenic influenza viruses.

概括 季节性流感病毒每年导致> 6亿例病例和数千亿美元 在损失中。大流行和禽流感病毒构成更大的威胁，因为它们可以 失调的免疫反应和/或传播到并关闭多个器官。当前的 有证据表明，异常病毒复制导致先天性的失调 免疫反应和低致病性高度致病菌株的出现 前体。 从低致病性病毒的过渡涉及插入多个碱性氨基 病毒血凝素（HA）表面蛋白的裂解位点的酸。在鸟类中，这种变化 HA允许该病毒系统地扩散，导致家禽中的死亡率高达100％。 氨基酸在裂解位点插入的分子机制不是很好 理解，但它可能涉及HA基因中RNA聚合酶的口吃，导致 核苷酸插入。 病毒RNA聚合酶还可以从病毒基因组中删除核苷酸，从而缩短 异常RNA。最近的研究表明，大流行和禽流感染了病毒感染 产生约56-125个核苷酸的RNA分子，长度称为迷你病毒RNA，并且 它们的合成与疾病标志物的上调相关。 RNA聚合酶如何 在病毒基因组中使如此大的缺失尚不清楚。 该项目将使用一种新颖的方法在病毒的关键步骤中停止病毒RNA聚合酶 复制和异常RNA合成，并使用最先进的生化，生物物理和 结构方法以揭示病毒复制和基因组封装的步骤 作为异常的RNA合成。这样，这个项目将有助于完整的机械 了解流感复制并回答有关长期存在的基本问题 高度致病的流感病毒的出现。