The biology of the nuclear export protein in influenza A virus replication

甲型流感病毒复制中核输出蛋白的生物学

• 批准号: 9975684
• 负责人: Benjamin R. tenOever
• 金额: $ 42.24万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-08 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975684
• 关键词: Antiviral Agents; Biology; Cell Culture Techniques; Cells; Complementary RNA; Complex; Data; Disease; Drug Design; Epitopes; Future; Genetic Transcription; Genome; Grant; Human; Immunoprecipitation; Infection; Influenza A virus; Integration Host Factors; Kinetics; Maps; Mass Spectrum Analysis; Mediating; Messenger RNA; Minor; Molecular; Molecular Target; Nature; Nuclear Export; Open Reading Frames; Pathogenicity; Play; Polymerase; Production; Protein Export Pathway; Proteins; RNA; RNA Processing; RNA Splicing; RNA Viruses; RNA chemical synthesis; RNA-Directed RNA Polymerase; Ribonucleoproteins; Role; Site; Species Specificity; Spliceosome Assembly Pathway; Transcript; Tropism; Variant; Viral; Viral Physiology; Viral Proteins; Virus; Virus Assembly; Virus Diseases; Virus Replication; base; bronchial epithelium; defined contribution; fitness; in vivo; mutant; next generation sequencing; novel; pathogenic virus; protein function; viral RNA

PROJECT SUMMARY In our continued efforts to understand the biology of Influenza A virus (IAV), we have characterized RNA from both the host and virus using next generation sequencing. These efforts led to the discovery of a small virus- derived RNA (svRNA) that was necessary for mediating the viral switch from transcription to replication. Upon characterizing the biology of svRNA, we documented the surprising finding that svRNA synthesis was dependent on the expression of NEP, a minor protein of the virus that slowly accumulates in the cell as a result of inefficient splicing. Together, these two discoveries suggested NEP may be the master regulator of IAV infection. In short, we found that the slow accumulation of NEP provided the virus with a “timer” to coordinate svRNA production and the subsequent switch from transcription to replication. Furthermore, we, and others, have found that NEP-mediated svRNA synthesis is dependent on its capacity to induce cRNA. Despite determining this function for NEP, how it coordinates this activity remains entirely unknown. Here we seek to understand this activity through three complementary aims. Aim 1 details a strategy to ascertain how NEP interaction with the RNA dependent RNA polymerase influences its activity. Aim 2 seeks to ascertain why NEP is associating with host factors involved in RNA processing. Aim 3 investigates the impact of NEP and its associated proteins on virus pathogenicity and tropism. The experimental strategy comprising these aims will reveal exciting new molecular targets that can be exploited to generate a novel class of antivirals and will significantly increase our understanding of IAV replication.

项目概要 在我们不断努力了解甲型流感病毒 (IAV) 的生物学过程中，我们对来自 A 型流感病毒 (IAV) 的 RNA 进行了表征 宿主和病毒均使用下一代测序。这些努力导致了一种小病毒的发现—— 衍生RNA（svRNA）是介导病毒从转录到复制的转变所必需的。之上 在描述 svRNA 的生物学特征时，我们记录了一个令人惊讶的发现，即 svRNA 的合成是 依赖于 NEP 的表达，NEP 是病毒的一种次要蛋白，因此会在细胞中缓慢积累 低效拼接。这两项发现共同表明 NEP 可能是 IAV 的主要调节者 感染。总之，我们发现NEP的缓慢积累为病毒提供了一个“计时器”来协调 svRNA 的产生以及随后从转录到复制的转换。此外，我们和其他人， 发现NEP介导的svRNA合成依赖于其诱导cRNA的能力。尽管 尽管确定了 NEP 的这一功能，但它如何协调这一活动仍然完全未知。在这里我们寻求 通过三个互补的目标来理解这项活动。目标 1 详细说明了确定 NEP 如何实施的策略 与RNA依赖性RNA聚合酶的相互作用影响其活性。目标 2 试图确定 NEP 的原因 与参与 RNA 加工的宿主因子相关。目标 3 调查 NEP 及其影响 病毒致病性和向性的相关蛋白。包含这些目标的实验策略将 揭示令人兴奋的新分子靶点，可用于产生一类新型抗病毒药物，并将 显着增加了我们对 IAV 复制的理解。