CRYSTAL STRUCTURE OF STRUCTURAL AND FUNCTIONAL PROTEINS FROM VESICULAR STOMATITI

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 水疱性口炎病毒(VSV)是一种非节段性负链RNA病毒，属于弹状病毒家族。VSV的11,161个核苷酸基因组包含5个基因，并被病毒核衣壳蛋白(N)所包裹。我们的团队已经开发了一种在大肠杆菌中共表达的系统，用于生产可溶性N蛋白，其功能取决于其与RNA结合并与病毒磷酸蛋白(P)形成复合体的能力。我们已经提纯并随后结晶了一种N蛋白低聚物(10-聚体)与90个核苷酸的RNA形成的复合体。此外，我们还将这一N蛋白-RNA低聚体与P蛋白的N蛋白结合域(P4)形成了络合物。与该项目一致的是，P蛋白的其他片段也已经结晶。已经收集了这些蛋白质和蛋白质复合体的初步数据集。N蛋白质-RNA复合体的模型正在建立中。目前，该病毒家族的N蛋白还没有高分辨率结构。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Vesicular stomatitis virus (VSV) is a non-segmented negative-stranded RNA virus belonging to the rhabdovirus family. The 11,161 nucleotide genome of VSV contains five genes and is encapsidated by the viral nucleocapsid protein(N). Our group has developed a co-expression system in E. coli for production of soluble N protein that is functional as determined by its abilty to bind RNA and form complexes with the viral phosphoprotein(P). We have purified and subsequently crystallized an N protein oligomer(a 10-mer) in complex with a 90 nucleotide RNA. In addition, we have crystallized this N protein-RNA oligomer in complex with the N protein binding domain of the P protein(P4). In line with the project, other fragments of the P protein have also been crystallized. Preliminary datasets for these proteins and protein complexes have been collected. Model building for the N protein-RNA complex is underway. Currently, there are no high resolution structures for the N proteins of this viral family.