CRYSTAL STRUCTURE OF STRUCTURAL AND FUNCTIONAL PROTEINS FROM VESICULAR STOMATITI

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 水泡性口炎病毒（VSV）是属于弹状病毒科的非节段负链RNA病毒。VSV的11，161个核苷酸的基因组包含5个基因，并被病毒核衣壳蛋白（N）包裹。本课题组成功地构建了大肠杆菌共表达系统。大肠杆菌中用于生产可溶性N蛋白，该可溶性N蛋白的功能性取决于其结合RNA并与病毒磷蛋白（P）形成复合物的能力。我们已经纯化并随后结晶了与90个核苷酸的RNA复合的N蛋白寡聚体（10聚体）。此外，我们已经结晶了这个N蛋白-RNA寡聚体与P蛋白（P4）的N蛋白结合结构域的复合物。根据该项目，P蛋白的其他片段也被结晶化。已经收集了这些蛋白质和蛋白质复合物的初步数据集。N蛋白-RNA复合物的模型构建正在进行中。目前，还没有高分辨率的结构为N蛋白的病毒家族。

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.