Probing the translational dynamics of influenza virus infection.

在细胞层面上，病毒感染的结果取决于病毒和细胞之间为控制细胞生物合成机制而进行的斗争，因为这最终决定了后代病毒的产量。许多病毒编码它们自己的核酸聚合酶，但它们都依赖于细胞蛋白质合成机制。因此，核糖体是关键战场。以人类和动物的主要病原体甲型流感病毒(IAV)作为模型系统，我们将使用核糖体图谱的新技术来详细说明这一冲突，这是迄今为止不可能的。核糖体图谱可以准确地确定核糖体在特定时刻翻译的是哪些mRNAs(病毒或细胞)，从而确定在争夺细胞控制权的战斗中“前线”的状态。我们的目标是：1.定义翻译成病毒-宿主细胞组合的细胞和病毒mRNAs，无论是病毒‘赢’还是宿主‘赢’。确定影响细胞基因表达的多种病毒机制及其特定靶点，这些机制决定了细胞翻译机器控制权之争的结果。测试IAV专门操纵核糖体使其偏向病毒信使优先翻译的假设。总体而言，该项目将从病毒和细胞角度定义关键事件，这些事件定义单个感染细胞是否被有效感染；数据将有助于设计新的治疗干预策略。

At the cellular level, the outcome of virus infection hinges on a battle between virus and cell for the control of the cellular biosynthetic machinery, as this ultimately determines the amount of progeny virus produced. Many viruses encode their own nucleic acid polymerases, but all are reliant on the cellular protein synthesis machinery. The ribosome therefore represents the key battleground. Using the major human and animal pathogen influenza A virus (IAV) as a model system, we will use the novel technique of ribosomal profiling to detail this conflict at a level of detail hitherto impossible. Ribosomal profiling allows the accurate determination of which mRNAs (viral or cellular) that the ribosomes are translating at a given moment; thereby defining the state of the 'frontline' in the battle for control of the cell. Our objectives are to:1. Define the cellular and viral mRNAs that are translated in virus-host cell combinations where either the virus 'wins' or where the host wins.2. To define which of the many viral mechanisms that affect cellular gene expression are and their specific targets which decide the outcome of the battle for control of the cellular translation machinery.3. Test the hypothesis that IAV specifically manipulates the ribosome to bias it towards preferential translation of viral messengers.Overall, the project will define the key events from both virus and cellular perspective that define whether an individual infected cell is productively infected; data that will aid the design of new therapeutic intervention strategies.