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.

项目总结 在我们不断努力了解甲型流感病毒(IAV)生物学的过程中，我们已经鉴定了来自 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型病毒-- 衍生的RNA(SvRNA)，是调节病毒从转录到复制的过程中所必需的。vt.在.的基础上 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：svRNA的合成是 依赖于NEP的表达，NEP是病毒的一种次要蛋白，因此在细胞中缓慢积累 低效的拼接。这两个发现表明，NEP可能是IAV的主要调节因子 感染。简而言之，我们发现NEP的缓慢积累为病毒提供了一个协调的“定时器” 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， 已发现NEP介导的svRNA合成依赖于其诱导cRNA的能力。尽管 确定NEP的这一功能，它如何协调这一活动仍然完全未知。在这里，我们试图 通过三个相辅相成的目标来理解这项活动。AIM 1详细介绍了一种确定NEP如何 与依赖于RNA的RNA聚合酶的相互作用会影响其活性。目标2试图确定为什么NEP 与RNA加工中涉及的宿主因素有关。目标3调查NEP的影响及其 病毒致病性和嗜性的相关蛋白。包含这些目标的试验性战略将 揭示令人兴奋的新分子靶点，可以用来产生一类新的抗病毒药物，并将 大大增加了我们对IAV复制的理解。