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

基本信息

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
项目类别：
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

项目摘要

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 进行了表征使用下一代测序对宿主和病毒进行研究，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。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这两项发现共同表明 NEP 可能是 IAV 的主要调节因子。总之，我们发现NEP的缓慢积累为病毒提供了一个协调的“计时器”。 svRNA 的产生以及随后从转录到复制的转换。发现NEP介导的svRNA合成依赖于其诱导cRNA的能力。确定 NEP 的这一功能，它如何协调这一活动仍然完全未知。这项活动通过三个互补的目标详细了解如何确定 NEP 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。目标 3 与参与 RNA 加工的宿主因素相关联，研究 NEP 及其影响。包含这些目标的实验策略将包括相关蛋白质对病毒致病性和趋向性的影响。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。