中文摘要
病毒离子通道蛋白可在宿主细胞膜上形成选择性离子通道,在病毒复制、装配和释放等过程中具有重要功能。现已证实黄病毒科的丙肝病毒(HCV)p7蛋白为离子通道蛋白,但同属黄病毒科的牛病毒性腹泻病毒(BVDV)离子通道蛋白的研究还未见报道。课题组前期的生物信息学分析和膜蛋白检测实验表明BVDV p7蛋白与HCV p7蛋白结构极其相似,BVDV p7为细胞膜蛋白,而且病毒离子通道蛋白抑制剂可抑制BVDV增殖。这些前期实验提示:BVDV p7可能是病毒离子通道蛋白且对BVDV的增殖有影响。本项目拟通过研究BVDV p7蛋白的亚细胞定位,寡聚体形成、p7对细胞膜通透性的影响等特性,明确BVDV p7蛋白的离子通道属性;利用反向遗传学技术构建p7突变的BVDV感染性cDNA克隆,研究p7蛋白在BVDV复制、装配和释放中的作用,为深入研究BVDV增殖机制、研发抗BVDV的药物和新型疫苗奠定基础。
英文摘要
Viral viroporin can form selective ion channel on the membrane of host cell, which has an important function for viral replication, assembly and release. Some studies have shown that p7 protein from flaviviridae hepatitis C virus (HCV) is a viroporin, but viroporin from bovine viral diarrhea virus (BVDV), as a member of the flaviviridae, have not been reported. Our bioinformatics studies display that the predicted structure of BVDV p7 protein is extremely similar to that of HCV p7 protein, which was furher confirmed by membrane protein analysis. Moreover, viroporin inhibitors can inhibit BVDV proliferation. These preliminary results suggest that BVDV p7 protein may be a viral viroporin and it have impact on the proliferation of BVDV. In this project, we will study BVDV p7 protein by subcellular localization, oligomer formation, p7 effects on membrane permeability and so on, which will confirm BVDV p7 is a viral viroporin or not; We will use reverse genetics technology to construct BVDV virion with p7 mutation, which will be used for studying the role of p7 protein in BVDV replication, assembly and release. Our results will provide a foundation for further studying BVDV mechanism, developing new drugs and anti-BVDV vaccines.
