Cassetti M C, Noah D L, Montelione G T, Krug R M
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey 08854, USA.
Virology. 2001 Oct 25;289(2):180-5. doi: 10.1006/viro.2001.1149.
We test the hypothesis that the translation machinery in cells infected by influenza A virus efficiently translates only mRNAs that possess the influenza viral 5' untranslated region (5'-UTR) by introducing mRNAs directly into the cytoplasm of infected cells. This strategy avoids effects due to the inhibition of the nuclear export of cellular mRNAs mediated by the viral NS1 protein. In one approach, we transfect in vitro synthesized mRNAs into infected cells and demonstrate that these mRNAs are efficiently translated whether or not they possess the influenza viral 5'-UTR. In the second approach, an mRNA is synthesized endogenously in the cytoplasm of influenza A virus infected cells by a constitutively expressed T7 RNA polymerase. Although this mRNA is uncapped and lacks the influenza viral 5'-UTR sequence, it is efficiently translated in infected cells via an internal ribosome entry site. We conclude that the translation machinery in influenza A virus infected cells is capable of efficiently translating all mRNAs and that the switch from cellular to virus-specific protein synthesis that occurs during infection results from other processes.
我们通过将mRNA直接导入受感染细胞的细胞质来检验这一假设:甲型流感病毒感染的细胞中的翻译机制仅能有效翻译具有流感病毒5'非翻译区(5'-UTR)的mRNA。该策略避免了由病毒NS1蛋白介导的细胞mRNA核输出抑制所产生的影响。在一种方法中,我们将体外合成的mRNA转染到受感染细胞中,并证明无论这些mRNA是否具有流感病毒5'-UTR,它们都能被有效翻译。在第二种方法中,一种mRNA由持续表达的T7 RNA聚合酶在甲型流感病毒感染细胞的细胞质中内源性合成。尽管这种mRNA没有帽结构且缺乏流感病毒5'-UTR序列,但它通过内部核糖体进入位点在受感染细胞中被有效翻译。我们得出结论,甲型流感病毒感染细胞中的翻译机制能够有效翻译所有mRNA,并且感染期间从细胞蛋白合成向病毒特异性蛋白合成的转变是由其他过程导致的。