Shen Shuo, Lin Pi-Shiu, Chao Yu-Chan, Zhang Aihua, Yang Xiaoming, Lim Seng Gee, Hong Wanjin, Tan Yee-Joo
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673, Singapore.
Biochem Biophys Res Commun. 2005 Apr 29;330(1):286-92. doi: 10.1016/j.bbrc.2005.02.153.
The severe acute respiratory syndrome coronavirus (SARS-CoV) 3a protein is one of the opening reading frames in the viral genome with no homologue in other known coronaviruses. Expression of the 3a protein has been demonstrated during both in vitro and in vivo infection. Here we present biochemical data to show that 3a is a novel coronavirus structural protein. 3a was detected in virions purified from SARS-CoV infected Vero E6 cells although two truncated products were present predominantly instead of the full-length protein. In Vero E6 cells transiently transfected with a cDNA construct for expressing 3a, a similar cleavage was observed. Furthermore, co-expression of 3a, membrane and envelope proteins using the baculovirus system showed that both full-length and truncated 3a can be assembled into virus-like particles. This is the first report that demonstrated the incorporation of 3a into virion and showed that the SARS-CoV encodes a novel coronavirus structural protein.
严重急性呼吸综合征冠状病毒(SARS-CoV)的3a蛋白是病毒基因组中的开放阅读框之一,在其他已知冠状病毒中没有同源物。3a蛋白在体外和体内感染过程中均有表达。在此,我们提供生化数据表明3a是一种新型冠状病毒结构蛋白。在从感染SARS-CoV的Vero E6细胞中纯化的病毒粒子中检测到了3a,不过主要存在的是两种截短产物而非全长蛋白。在用表达3a的cDNA构建体瞬时转染的Vero E6细胞中,也观察到了类似的切割现象。此外,使用杆状病毒系统共表达3a、膜蛋白和包膜蛋白表明,全长和截短的3a都能组装成病毒样颗粒。这是首次证明3a整合到病毒粒子中,并表明SARS-CoV编码一种新型冠状病毒结构蛋白的报告。