Updike W S, Tesar M, Ehrenfeld E
Department of Cellular, Viral, and Molecular Biology, University of Utah School of Medicine, Salt Lake City 84132.
Virology. 1991 Nov;185(1):411-8. doi: 10.1016/0042-6822(91)90789-e.
Hepatitis A virus (HAV) is distinguished from other picornaviruses by its tropism for the liver in infected hosts, a nonlytic infection in hepatocytes, and a slow and nonlytic growth cycle in cultured cells. Although the genome structure and organization of HAV appear to be similar to those of the other picornaviruses, the viral proteins synthesized in infected cells have not been previously characterized. We have utilized specific antisera raised in rabbits to recombinant HAV proteins expressed in Escherichia coli in an effort to identify both structural and nonstructural proteins in BS-C-1 cells throughout the course of a viral replication cycle. Replication was monitored by dot blot hybridization of viral genomes. Structural proteins VP0, VP1, VP2, and VP3 were found to accumulate during the infection cycle as did viral RNA. Nonstructural proteins 2C and 3D were not detected on immunoblots, although a minor amount of 2C could be detected by immunoprecipitation of lysates of radiolabeled, infected cells. The relative sensitivities of the various antisera were determined, and the failure to observe nonstructural proteins was shown not to be due to decreased sensitivity of the detection reagents. Thus, it appears that HAV nonstructural proteins do not accumulate in infected cells to levels comparable to those of capsid proteins.
甲型肝炎病毒(HAV)与其他小核糖核酸病毒的区别在于,它在受感染宿主中对肝脏具有嗜性,在肝细胞中呈非溶细胞性感染,并且在培养细胞中具有缓慢且非溶细胞性的生长周期。尽管HAV的基因组结构和组织似乎与其他小核糖核酸病毒相似,但此前尚未对受感染细胞中合成的病毒蛋白进行表征。我们利用在兔子体内产生的针对在大肠杆菌中表达的重组HAV蛋白的特异性抗血清,试图在病毒复制周期的全过程中鉴定BS-C-1细胞中的结构蛋白和非结构蛋白。通过病毒基因组的斑点杂交监测复制情况。发现结构蛋白VP0、VP1、VP2和VP3在感染周期中与病毒RNA一样积累。在免疫印迹上未检测到非结构蛋白2C和3D,尽管通过对放射性标记的受感染细胞裂解物进行免疫沉淀可检测到少量的2C。确定了各种抗血清的相对敏感性,并且表明未观察到非结构蛋白并非由于检测试剂的敏感性降低。因此,似乎HAV非结构蛋白在受感染细胞中的积累水平与衣壳蛋白不同。