PICORNAVIRUS-HUMAN PROTEIN INTERACTION ANALYSIS BY PROTEIN AFFINITY AND MS/MS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目的重点是了解人类和小核糖核酸病毒蛋白之间的相互作用。 小核糖核酸病毒是无包膜、单链、正极性RNA病毒，其引起许多重要的人类和动物疾病，包括脊髓灰质炎、甲型肝炎、口蹄疫和普通感冒（鼻病毒）。 虽然许多工作已经进入了解脊髓灰质炎的六个非结构基因的复制和宿主翻译关闭机制的目标，很少有人知道其他小核糖核酸病毒的结合靶点。 我们特别感兴趣的是新的小核糖核酸病毒，如克拉塞病毒，并比较宿主病毒蛋白质的相互作用，在这个系统中更好地表征小核糖核酸病毒，如脊髓灰质炎，以及小核糖核酸病毒家族，如甲型肝炎和柯萨奇病毒的更不同的成员。 为此，我们使用亲和纯化在人293细胞中表达的病毒蛋白来捕获蛋白相互作用复合物，然后进行LC-MS/MS分析。 这些工作有助于解释小核糖核酸病毒家族不同成员之间在毒力、复制、翻译和嗜性方面的差异。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is focused on understanding the interaction between human and picornaviral proteins. Picornaviruses are non-enveloped, single-stranded, positive polarity RNA viruses that cause many important human and animal diseases, including polio, hepatitis A, foot-and-mouth disease, and the common cold (rhinovirus). Although much work has gone into understanding the targets of the six nonstructural genes of polio in regards to the mechanism of replication and host-translational shut-off, very little is known about the binding targets of other picornaviruses. We are particularly interested in novel picornaviruses such as klassevirus, and are comparing host-virus protein interactions in this system with better characterized picornaviruses such as polio, as well as more divergent members of the picornavirus family such as Hepatitis A, and Coxsackievirus. To this end, we are using affinity purification of virus proteins expressed in human 293 cells to capture protein-interaction complexes, followed by LC-MS/MS analysis. Such work could help explain difference in virulence, replication, translation, and tropism between divergent members of the picornavirus family.