Mechanism for Intercellular Movement of Potato Virus X

PVX的三重基因块是一个遗传模块，在Potex-、Carla-、Furo-和HordeiVirus家族成员中保守，是病毒通过胞间连丝传播所必需的。PVX三重基因块产生三种蛋白质，分别为25K、12K和8K蛋白质。虽然这些蛋白质的几个性质已经确定，但这些蛋白质在运动过程中的确切贡献还没有确定。该项目旨在确定PVX蛋白的特定功能，分析它们如何相互作用以促进病毒移动，并确定它们如何与宿主细胞相互作用以促进病毒移动。制备了含有PVX运动蛋白基因和GFP基因融合的质粒，并用于轰击烟草叶片。用荧光显微镜观察蛋白质在细胞间的运动。将质粒轰击到表达PVX三基因阻断蛋白的转基因烟草上，以研究病毒蛋白-蛋白相互作用对病毒蛋白运动和亚细胞靶标的影响。这种方法非常有价值，因为它提供了一个机会来观察一些以前在利用显微注射将蛋白质输送到细胞中的研究中没有发现的蛋白质-蛋白质相互作用。其次，将使用PVX 25K蛋白进行突变分析，以确定在蛋白质运动和蛋白质之间相互作用中起作用的结构域。这些研究将与分析体内病毒粒子组装和确定病毒粒子在细胞间运动中的作用的创新方法相结合。

The triple gene block of PVX is a genetic module that is conserved among members of the potex-, carla-, furo-, and hordeivirus families and is required for virus transport through plasmodesmata. The PVX triple gene block produces three proteins named the 25K, 12K and 8K proteins. While several properties of these proteins have been identified the exact contribution of these proteins to the movement process has not been determined yet. The project is designed to identify the specific functions of the PVX proteins, to analyze how they interact with each other to promote virus movement, and to determine how they interact with the host cell to facilitate virus movement. Plasmids containing the PVX movement protein genes fused to the GFP gene were prepared and used to bombard tobacco leaves. Movement of proteins between cells is viewed by fluorescence microscopy. Plasmids will be bombarded to transgenic tobacco expressing the PVX triple gene block proteins to study the effects of viral protein-protein interactions on viral protein movement and subcellular targetting. This approach has been quite valuable because it has provided an opportunity to view some protein-protein interactions that were not detected previously in studies which utilized microinjection to deliver proteins into cells. Secondly, mutational analysis will be conducted using the PVX 25K protein to identify domains that function in protein movement and protein-protein interactions. These studies will be combined with innovative approaches to analyze virion assembly in vivo and to determine the role of virions in cell-to-cell movement.