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.

项目摘要 在我们继续努力了解流感病毒（IAV）的生物学的努力中，我们表征了RNA 宿主和病毒都使用下一代测序。这些努力导致发现了一种小病毒 - 衍生的RNA（SVRNA）是介导从转录到复制的病毒切换所必需的。之上 表征SVRNA的生物学，我们记录了Svrna合成的令人惊讶的发现 依赖于NEP的表达，NEP的表达是一种较小的病毒蛋白，结果慢慢积聚在细胞中 效率低下的剪接。这两个发现共同表明NEP可能是IAV的主要调节剂 感染。简而言之，我们发现NEP的缓慢积累为病毒提供了“计时器”以协调 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， 已经发现NEP介导的SVRNA合成取决于其诱导CRNA的能力。尽管 确定NEP的此功能，它如何协调此活动仍然完全未知。我们在这里寻求 通过三个完整的目标了解这项活动。目标1详细介绍了确定如何NEP的策略 与RNA依赖性RNA聚合酶的相互作用会影响其活性。 AIM 2试图确定为什么NEP 正在与RNA处理中涉及的宿主因素相关联。 AIM 3调查了NEP及其的影响 相关蛋白质在病毒的致病性和向性方面。完成这些目标的实验策略将 揭示令人兴奋的新分子靶标，可以探索以生成新型的抗病毒药物，并将 显着增加了我们对IAV复制的理解。