Protein Interactions During Replication of a Plant Rhabodovirus

;​R = E = E = F = F；C o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o ol;8 9:；= ?Microsoft Word 6.0文档MSWordDoc。6;​本研究旨在研究植物横纹肌病毒（sonchus yellow net virus， SYNV）的复制和包装过程中病毒蛋白的细胞内定位，以及该病毒在植物细胞核内复制和随后从核膜出芽过程中发生的生物化学事件。这是世界上为数不多的实验室之一在细胞和分子水平上研究植物中这类重要的包膜病毒的持续研究。SYNV的生物学挑战了动物横纹肌病毒生物学的范式。与动物横纹肌病毒，水疱性口炎病毒（VSV）不同，它在细胞质和细胞膜芽中复制，从而作为分泌膜运输的模型，SYNV在核膜的细胞核和芽中复制。该提案将研究这些差异，包括检查在动物横纹肌病毒中未发现的基因产物sc4的性质。该产物缺乏跨膜结构域，其核靶向性特别令人感兴趣。其他蛋白质的核靶向结构域的性质是有兴趣的，以便为该系统奠定坚实的基础。由于复制发生在细胞核中，因此将通过分析转录活性蛋白的蛋白质-蛋白质相互作用以及纯化病毒的转录无活性核心颗粒与转录活性核复合物的比较，探索SYNV复制复合物的潜在新方面。与动物病毒相比，关于植物横纹肌病毒的信息是有限的。然而，正如其他既包含动物病毒又包含植物病毒的类群（如小核糖核酸和辛德比病毒类病毒类群以及布尼亚病毒科）的案例所显示的那样，对这两个类群的平行成员的研究往往使我们对这些病毒复制的理解有了更快的发展。将这种病毒的成分靶向进入细胞核的研究也是及时的，因为将选定的蛋白质靶向进入细胞核腔和将蛋白质整合到植物和动物细胞的核膜的机制尚不清楚。这里有潜力开发模型蛋白来检查瞄准核膜的目标，这是一个知之甚少的领域。植物横纹肌病毒也可作为检测植物核/细胞质中蛋白质流动和含病毒粒子囊泡胞内流动的一种有价值的系统。*** ;0 + 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0$ H l D H R:\WWUSER\TEMPLATE\NORMAL。DOT Lawrence R. Griffing Una Solomon @ g @ @ Microsoft Word 6.0E = E p p p p p pC 1 Õ % t t 3 C p p p p Õ p p p p p pÕ 7 9418086 Jackson这个建议解决了细胞内的损失

; R o o t E n t r y F l; C o m p O b j b W o r d D o c u m e n t O b j e c t P o o l l; l; 8 9 : ; = ? @ A F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; 9418086 Jackson This proposal addresses the intracellular location of viral proteins involved in replication and packaging the plant rhabdovirus, sonchus yellow net virus (SYNV), and the biochemistry of the events occurring during replication of the virus in the plant nucleus and subsequent budding from the nuclear membrane. This is a continuing study from one of the few labs in the world studying this important class of enveloped virus in plants at the cellular and molecular level. The biology of SYNV challenges the paradigms of animal rhabdovirus biology. Unlike the animal rhabdovirus, Vesicular Stomatitis Virus (VSV), which replicates in cytoplasm and buds from the cell membrane, thereby serving as a model for secretory membrane traffic, SYNV replicates in the nucleus and buds from the nuclear membrane . This proposal will study these differences, including examining the nature of a gene product, sc4, not found in animal rhabdoviruses. The nuclear targeting of this product, which lacks a transmembrane domain, is of particular interest. The nature of the nuclear targeting domains of th e other proteins is of interest in order to lay a firm groundwork for this system. Since replication occurs in the nucleus, potentially novel aspects of the replication complex of SYNV will be explored through the analysis of protein-protein interaction of transcription-active proteins and the comparison of the transcriptionally inactive core particles from purified virus with transcriptionally active nuclear complexes. %%% Information on plant Rhabdovirusues is limited in comparison to their animal counterparts. Yet, as has been shown in the case of other groups that encompass both animal and plant viruses, such as the picorna- and sindbis-like virus groups, and the Bunyaviridae, the study in parallel members of both kingdoms has often led to a much more rapid development of our understanding of replication of these viruses. Investigations into targeting the components of this virus into the nucleus are also timely because the mechanism of targeting select proteins into the lumen of the nucleus and of integration of proteins into the nuclear membrane of plant and animal cells is unclear. There is the potential here for developing model proteins to examine targeting to the inner nuclear envelope, an area where there is very little knowledge. The plant rhabdoviruses could also provide a valuable system to examine nuclear/cytoplasmic flow of protein and the intracellular flow of virion-containing vesicles in plants. *** ; Oh +' 0 S u m m a r y I n f o r m a t i o n ( 7 $ H l D h R:\WWUSER\TEMPLATE\NORMAL.DOT Lawrence R. Griffing Una Solomon @ g @ @ @ Microsoft Word 6.0 4 e = e p p p p p p p C 1 Õ % t T 3 C p C p p p p Õ p p p p Õ 7 9418086 Jackson This proposal addresses the intracellular lo