Rhabdovirus phosphoproteins: RNA silencing and complex formation

产品说明：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水稻黄矮病毒。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。拟议的实验将测试的假设，磷酸化的“P”蛋白编码的植物负链RNA病毒具有新的功能，相对于同源蛋白从动物病毒，这是所需的成功感染植物细胞。我的实验室已经提供了实验证据，扩展了P的多种活性，包括RNA沉默抑制因子（RSS）。我们还发现，这种活性可以通过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这种活性如何通过与其他SYNV蛋白的相互作用来调节。具体目的1：检验SYNV-P的RSS活性是通过直接结合RNA介导的假设。具体目的2：检验RSS活性在弹状病毒P蛋白中是保守的这一假设。具体目标3：为了检验SYNV-P的RSS活性可以通过与核衣壳（N）或基质（M）蛋白的相互作用来调节的假设。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肯塔基州大学NIH资助的项目，该项目可以确定这些蛋白质的X射线晶体结构。迄今为止，由弹状病毒编码的N和P蛋白质的结构与动物宿主尚未确定。因此，如果成功的话，这些蛋白质中的任何一种在同质或异质复合物中的结晶都将代表来自不同学科的大量研究人员所重视的重要贡献。总之，这项研究对于基本了解弹状病毒的分子和细胞生物学，特别是植物弹状病毒的分子和细胞生物学具有重要意义，并有可能导致开发各种疾病的新控制策略。

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.