Casais Rosa, Davies Marc, Cavanagh David, Britton Paul
Division of Molecular Biology, Institute for Animal Health, Compton Laboratory, Compton, Newbury, Berkshire, RG20 7NN, United Kingdom.
J Virol. 2005 Jul;79(13):8065-78. doi: 10.1128/JVI.79.13.8065-8078.2005.
The avian coronavirus Infectious bronchitis virus (IBV), like other coronaviruses, expresses several small nonstructural (ns) proteins in addition to those from gene 1 (replicase) and the structural proteins. These coronavirus ns genes differ both in number and in amino acid similarity between the coronavirus groups but show some concordance within a group or subgroup. The functions and requirements of the small ns gene products remain to be elucidated. With the advent of reverse genetics for coronaviruses, the first steps in elucidating their role can be investigated. We have used our reverse genetics system for IBV (R. Casais, V. Thiel, S. G. Siddell, D. Cavanagh, and P. Britton, J. Virol. 75:12359-12369, 2001) to investigate the requirement of IBV gene 5 for replication in vivo, in ovo, and ex vivo. We produced a series of recombinant viruses, with an isogenic background, in which complete expression of gene 5 products was prevented by the inactivation of gene 5 following scrambling of the transcription-associated sequence, thereby preventing the expression of IBV subgenomic mRNA 5, or scrambling either separately or together of the translation initiation codons for the two gene 5 products. As all of the recombinant viruses replicated very similarly to the wild-type virus, Beau-R, we conclude that the IBV gene 5 products are not essential for IBV replication per se and that they are accessory proteins.
禽冠状病毒传染性支气管炎病毒(IBV)与其他冠状病毒一样,除了来自基因1(复制酶)的蛋白和结构蛋白外,还表达几种小的非结构(ns)蛋白。这些冠状病毒的ns基因在冠状病毒组之间的数量和氨基酸相似性方面都有所不同,但在一个组或亚组内表现出一定的一致性。小ns基因产物的功能和需求仍有待阐明。随着冠状病毒反向遗传学的出现,可以研究阐明它们作用的第一步。我们利用我们的IBV反向遗传学系统(R. Casais、V. Thiel、S. G. Siddell、D. Cavanagh和P. Britton,《病毒学杂志》75:12359 - 12369,2001年)来研究IBV基因5在体内、卵内和体外复制中的需求。我们构建了一系列具有同基因背景的重组病毒,其中通过转录相关序列混乱后基因5的失活来阻止基因5产物的完全表达,从而阻止IBV亚基因组mRNA 5的表达,或者分别或一起打乱两个基因5产物的翻译起始密码子。由于所有重组病毒的复制方式与野生型病毒Beau - R非常相似,我们得出结论,IBV基因5产物本身对于IBV复制不是必需的,它们是辅助蛋白。