Rhabdovirus phosphoproteins: RNA silencing and complex formation

弹状病毒磷蛋白：RNA 沉默和复合物形成

• 批准号: 7197074
• 负责人: Michael M Goodin
• 金额: $ 6.8万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7197074
• 关键词: Affinity; Animal Viruses; Animals; Binding; Cell Nucleus; Cells; Cellular biology; Communities; Complex; Crystallization; Data; Development; Discipline; Disease; Double-Stranded RNA; Electrophoretic Mobility Shift Assay; Family; Fishes; Funding; Future; Genes; Genetic Transcription; Glycine decarboxylase; Goals; Health; Human; In Vitro; Infection; Infectious hematopoietic necrosis virus; Insecta; Kentucky; Lasers; Lead; Length; Mediating; Methods; Microscope; Molecular; Nucleocapsid; Phosphoproteins; Plant Viruses; Plants; Play; Polymerase; Proteins; RNA; RNA Binding; RNA Interference; RNA Recognition Motif; RNA Viruses; RNA-Binding Proteins; Rabies virus; Reagent; Relative (related person); Replication-Associated Process; Research; Research Personnel; Rhabdoviridae; Rice; Roentgen Rays; Role; Scanning; Sonchus; Structure; Sum; System; Testing; United States National Institutes of Health; Universities; Viral; Viral Proteins; Virus; base; in vivo; insight; member; multiple myeloma M Protein; mutant; novel; pathogen; protein function; protein structure; research study; transcription factor; viral RNA

DESCRIPTION: The family Rhabdoviridae has members that represent some of the greatest threats to human, animal and plant health, including Rabies virus of humans and animals, Infectious hematopoietic necrosis virus offish, and Rice yellow stunt virus of plants. By comparing the functions of proteins encoded by different rhabdoviruses we hope to fulfill our long-term goal, which is to understand how new viral pathogens emerge via adaptive changes required for replication in different host cells. The proposed experiments will test the hypothesis that phosphorylated "P" proteins encoded by plant negative-strand RNA viruses have novel functions, relative to the cognate proteins from animal viruses, which are required for the successful infection of plant cells. My lab has provided experimental evidence that extends the multiple activities of P to include that of RNA silencing suppressor (RSS). We have also made the novel discovery that this activity can be regulated by its interaction with a second viral protein. The objective of this application is to elucidate the mechanism by which SYNV-P functions as an RSS and how this activity is moderated by interaction with other SYNV proteins. Specific Aim 1: To test the hypothesis that the RSS activity of SYNV-P is mediated via direct binding to RNA. Specific Aim 2: To test the hypothesis that RSS activity is conserved among rhabdovirus P proteins. Specific Aim 3: To test the hypothesis that the RSS activity of SYNV-P can be modulated via interactions with the nucleocapsid (N) or matrix (M) proteins. Additionally, our expertise in purifying N and P protein complexes from insect and bacterial cells will enable us to take advantage of an NIH-funded project at the University of Kentucky that could determine the X-ray crystal structures for these proteins. To date, the structures of the N and P proteins encoded by rhabdoviruses with animal hosts have not been determined. Therefore, if successful, the crystallization of any of these proteins, in homo- or heterogeneous complexes, would represent an important contribution valued by a large community of researchers from different disciplines. In sum, this research is significant for its contribution to the basic understanding of the molecular and cell biology of rhabdoviruses, particularly those of plants, and has the potential to lead to the development of novel control strategies for a variety of diseases.

描述：弹状病毒科的成员对人类、动物和植物健康构成最大威胁，包括人类和动物的狂犬病病毒、鱼类的传染性造血坏死病毒和植物的稻黄矮化病毒。通过比较不同弹状病毒编码的蛋白质的功能，我们希望实现我们的长期目标，即了解新的病毒病原体如何通过在不同宿主细胞中复制所需的适应性变化而出现。拟议的实验将验证以下假设：相对于动物病毒的同源蛋白，植物负链RNA病毒编码的磷酸化“P”蛋白具有新颖的功能，这是成功感染植物细胞所必需的。我的实验室提供了实验证据，将 P 的多种活性扩展到包括 RNA 沉默抑制子 (RSS) 的活性。我们还发现，这种活性可以通过其与第二种病毒蛋白的相互作用来调节。本申请的目的是阐明 SYNV-P 作为 RSS 发挥作用的机制，以及如何通过与其他 SYNV 蛋白的相互作用来调节这种活性。具体目标 1：检验 SYNV-P 的 RSS 活性是通过直接结合 RNA 介导的假设。具体目标 2：检验弹状病毒 P 蛋白中 RSS 活性保守的假设。具体目标 3：检验 SYNV-P 的 RSS 活性可通过与核衣壳 (N) 或基质 (M) 蛋白相互作用进行调节的假设。此外，我们在从昆虫和细菌细胞中纯化 N 和 P 蛋白复合物方面的专业知识将使我们能够利用 NIH 资助的肯塔基大学项目，该项目可以确定这些蛋白质的 X 射线晶体结构。迄今为止，动物宿主弹状病毒编码的N和P蛋白的结构尚未确定。因此，如果成功，这些蛋白质中的任何一种，无论是同源还是异源复合物的结晶，都将代表来自不同学科的众多研究人员所重视的重要贡献。总之，这项研究对于弹状病毒（尤其是植物弹状病毒）的分子和细胞生物学的基本理解做出了重要贡献，并且有可能导致开发针对多种疾病的新型控制策略。