Tylor Shaun, Andonov Anton, Cutts Todd, Cao Jingxin, Grudesky Elsey, Van Domselaar Gary, Li Xuguang, He Runtao
National Microbiology Laboratory, Health Canada, 1015 Arlington St, Winnipeg, MB R3E3R2, Canada.
Can J Microbiol. 2009 Mar;55(3):254-60. doi: 10.1139/w08-139.
The multimerization/self-interaction of viral proteins is an important step in the process of viral assembly and maturation. Our previous study indicated that the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid protein forms self-multimers through a serine-arginine (SR)-rich motif (SSRSSSRSRGNSR) by using a mammalian two-hybrid system. To determine the biological relevance of this motif, we constructed a SARS-CoV reverse genetic construct by using a bacterial artificial chromosome (BAC)-based vector controlled by a T7 promoter; and subsequently deleted the SR-rich motif from the N gene. The mutated infectious clone showed reduced level of genome transcription and significantly reduced levels of the infectious virions. These results strongly suggest that the SR-rich motif is critical for effective virus replication.
病毒蛋白的多聚化/自身相互作用是病毒组装和成熟过程中的重要步骤。我们之前的研究表明,严重急性呼吸综合征相关冠状病毒(SARS-CoV)核衣壳蛋白通过富含丝氨酸-精氨酸(SR)的基序(SSRSSSRSRGNSR)利用哺乳动物双杂交系统形成自身多聚体。为了确定该基序的生物学相关性,我们使用由T7启动子控制的基于细菌人工染色体(BAC)的载体构建了SARS-CoV反向遗传构建体;随后从N基因中删除了富含SR的基序。突变的感染性克隆显示基因组转录水平降低,感染性病毒粒子水平显著降低。这些结果强烈表明,富含SR的基序对有效的病毒复制至关重要。