Regulating Influenza Polymerase Structure and Function by Phosphorylation

通过磷酸化调节流感聚合酶的结构和功能

• 批准号: 8367844
• 负责人: Andrew Mehle
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-07 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8367844
• 关键词: Avian Influenza A Virus; Biochemical Genetics; Birds; Chromosome Mapping; Complex; DNA Sequence Rearrangement; Event; Foundations; Future; Gene Expression; Genome; Human; Infection; Influenza; Influenza A virus; Instruction; Integration Host Factors; Molecular; Nucleoproteins; Pathogenicity; Pathway interactions; Phosphorylation; Phosphorylation Site; Polymerase; Population; Process; Proteins; Regulation; Resolution; Structure; System; Therapeutic Intervention; Translations; Viral; Viral Proteins; Virus Replication; flu transmission; influenza outbreak; influenzavirus; insight; loss of function; prevent; protein function; transmission process; viral RNA

Protein function is dynamically controlled by post-translation events. A connmon regulatory mechanism is the phosphorylation and subsequent conformational rearrangement of target proteins. Viral proteins are often controlled by the same pathways that target proteins encoded by the host. The replication cycle of influenza A virus is influenced by host proteins and provides a unique system in which to study these processes. A major target of regulation is the influenza virus replication machinery containing viral RNA, nucleoprotein (NP), and the trimeric polymerase composed ofthe proteins PB1, PB2, and PA. The replication machinery controls the ordered transition from gene expression to genome replication that is essential for a productive infection. In addition, the viral polymerase is a key determinant in the host range of influenza virus and restricts the transmission of influenza virus from avian to human populations. The processes controlling the replication machinery and the interplay between the polymerase and the host are poorly understood. We propose an integrative approach using biochemical, genetic, and structural studies to determine the host proteins and molecular mechanisms that regulate the influenza replication machinery. Specifically, Aim 1 will identify sites of phosphorylation within viral proteins and examine the functional consequences of phosphorylation on virus replication and the determination of host range. Aim 2 will build on recent genetic mapping of interactions to produce high-resolution crystal structures that characterize in detail the protein interfaces of the replication complex. Finally, Aim 3 will exploit loss-of-function screens to identify cellular proteins that regulate the influenza polymerase and control transmission of influenza from birds to humans. These studies will provide crucial insight into the viral and host factors controlling influenza replication and will provide the foundation for rational strategies to treat and prevent future influenza outbreaks in humans.

蛋白质的功能受翻译后事件的动态控制。一个狡猾的监管者 其作用机制是靶蛋白的磷酸化和随后的构象重排。 病毒蛋白通常由与宿主编码的目标蛋白相同的途径控制。 甲型流感病毒的复制周期受宿主蛋白的影响，并提供独特的 研究这些过程的系统。监管的主要目标是流感病毒。 含有病毒RNA、核蛋白(NP)和三聚体聚合酶的复制机制 由蛋白质PB1、PB2和PA组成。复制机械控制着有序的 从基因表达到基因组复制的过渡，这对生产性感染是必不可少的。在……里面 此外，病毒聚合酶是流感病毒宿主范围内的关键决定因素，并限制 流感病毒从禽类传播到人类。控制进程的进程 复制机制以及聚合酶和宿主之间的相互作用还知之甚少。 我们提出了一种综合的方法，使用生化、遗传和结构研究来确定 调节流感复制机制的宿主蛋白质和分子机制。 具体地说，AIM 1将识别病毒蛋白中的磷酸化位点，并检查 磷酸化对病毒复制的功能影响和宿主范围的确定。 AIM 2将建立在最近相互作用的遗传图谱的基础上，以生产高分辨率晶体 详细表征复制复合体的蛋白质界面的结构。最后，目标3 将利用功能丧失筛查来识别调节流感聚合酶的细胞蛋白 并控制流感从禽类向人类的传播。这些研究将提供至关重要的见解 深入研究控制流感复制的病毒和宿主因素，并将为 治疗和预防人类未来流感暴发的合理策略。