Neumann Gabriele, Feldmann Heinz, Watanabe Shinji, Lukashevich Igor, Kawaoka Yoshihiro
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
J Virol. 2002 Jan;76(1):406-10. doi: 10.1128/jvi.76.1.406-410.2002.
Ebola virus, a prime example of an emerging pathogen, causes fatal hemorrhagic fever in humans and in nonhuman primates. Identification of major determinants of Ebola virus pathogenicity has been hampered by the lack of effective strategies for experimental mutagenesis. Here we exploit a reverse genetics system that allows the generation of Ebola virus from cloned cDNA to engineer a mutant Ebola virus with an altered furin recognition motif in the glycoprotein (GP). When expressed in cells, the GP of the wild type, but not of the mutant, virus was cleaved into GP1 and GP2. Although posttranslational furin-mediated cleavage of GP was thought to be an essential step in Ebola virus infection, generation of a viable mutant Ebola virus lacking a furin recognition motif in the GP cleavage site demonstrates that GP cleavage is not essential for replication of Ebola virus in cell culture.
埃博拉病毒是一种新出现病原体的典型例子，可在人类和非人类灵长类动物中引发致命的出血热。由于缺乏有效的实验诱变策略，埃博拉病毒致病性主要决定因素的鉴定工作受到了阻碍。在此，我们利用一种反向遗传学系统，该系统可从克隆的互补脱氧核糖核酸（cDNA）产生埃博拉病毒，从而构建出一种在糖蛋白（GP）中改变了弗林蛋白酶识别基序的突变型埃博拉病毒。当在细胞中表达时，野生型病毒而非突变型病毒的GP被切割成GP1和GP2。尽管翻译后弗林蛋白酶介导的GP切割被认为是埃博拉病毒感染的一个关键步骤，但构建出一种在GP切割位点缺乏弗林蛋白酶识别基序的存活突变型埃博拉病毒表明，GP切割对于埃博拉病毒在细胞培养中的复制并非必不可少。
