Zhou Bing, Liu Junli, Wang Qiuna, Liu Xuan, Li Xiaorong, Li Ping, Ma Qingjun, Cao Cheng
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, Beijing, China.
J Virol. 2008 Jul;82(14):6962-71. doi: 10.1128/JVI.00133-08. Epub 2008 Apr 30.
Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of SARS, an emerging disease characterized by atypical pneumonia. Using a yeast two-hybrid screen with the nucleocapsid (N) protein of SARS-CoV as a bait, the C terminus (amino acids 251 to 422) of the N protein was found to interact with human elongation factor 1-alpha (EF1alpha), an essential component of the translational machinery with an important role in cytokinesis, promoting the bundling of filamentous actin (F-actin). In vitro and in vivo interaction was then confirmed by immuno-coprecipitation, far-Western blotting, and surface plasmon resonance. It was demonstrated that the N protein of SARS-CoV induces aggregation of EF1alpha, inhibiting protein translation and cytokinesis by blocking F-actin bundling. Proliferation of human peripheral blood lymphocytes and other human cell lines was significantly inhibited by the infection of recombinant retrovirus expressing SARS-CoV N protein.
严重急性呼吸综合征冠状病毒(SARS-CoV)是SARS的病原体,SARS是一种以非典型肺炎为特征的新出现疾病。利用酵母双杂交筛选,以SARS-CoV的核衣壳(N)蛋白作为诱饵,发现N蛋白的C末端(氨基酸251至422)与人类延伸因子1-α(EF1α)相互作用,EF1α是翻译机制的一个重要组成部分,在胞质分裂中起重要作用,促进丝状肌动蛋白(F-肌动蛋白)的成束。然后通过免疫共沉淀、远缘Western印迹和表面等离子体共振证实了体外和体内的相互作用。结果表明,SARS-CoV的N蛋白诱导EF1α聚集,通过阻断F-肌动蛋白成束抑制蛋白质翻译和胞质分裂。表达SARS-CoV N蛋白的重组逆转录病毒感染显著抑制了人外周血淋巴细胞和其他人细胞系的增殖。