de la Luna S, Fortes P, Beloso A, Ortín J
Centro Nacional de Biotecnología (Consejo Superior de Investigaciones Cientifícas), Universidad Autónoma de Madrid, Spain.
J Virol. 1995 Apr;69(4):2427-33. doi: 10.1128/JVI.69.4.2427-2433.1995.
The effect of NS1 protein on the efficiency of influenza virus mRNA translation was evaluated by determining the accumulation of nucleoprotein (NP) or M1 mRNAs in the cytoplasm of cells expressing either of these genes alone or in combination with the NS1 gene, as well as the total cell accumulation of NP or M1 protein. Coexpression of NS1, but not of NS2 protein, led to increases in the translation of these mRNAs in the range of 5- to 100-fold. This translation enhancement was specific for viral mRNAs, since the translation of neither cat nor lacZ mRNAs was affected by the coexpression of NS1 protein. The use of chimeric cat genes containing the 5'-extracistronic sequences of the influenza virus mRNAs corresponding to segment 2, 7, or 8 indicated that these sequences can in part account for the observed effect. The enhancement of viral mRNA translation mediated by NS1 protein was due to an increase in the translation initiation rate, since the sizes of NP-specific polysomes, but not those of lacZ-specific polysomes, was significantly higher in cells coexpressing NS1 protein than in those expressing only the NP gene.
通过测定单独表达核蛋白(NP)或M1基因,或与NS1基因共同表达这些基因的细胞胞质中NP或M1 mRNA的积累情况,以及NP或M1蛋白在细胞中的总积累量,评估NS1蛋白对流感病毒mRNA翻译效率的影响。NS1(而非NS2蛋白)的共表达导致这些mRNA的翻译增加了5至100倍。这种翻译增强对病毒mRNA具有特异性,因为NS1蛋白的共表达对cat和lacZ mRNA的翻译均无影响。使用含有对应于第2、7或8节段的流感病毒mRNA 5'-顺反子外序列的嵌合cat基因表明这些序列可部分解释所观察到的效应。NS1蛋白介导的病毒mRNA翻译增强是由于翻译起始速率的增加,因为在共表达NS1蛋白的细胞中,NP特异性多核糖体的大小显著高于仅表达NP基因的细胞中的大小,而lacZ特异性多核糖体的大小则无显著差异。