Dhar D, Mapa K, Pudi R, Srinivasan P, Bodhinathan K, Das S
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, India.
Arch Virol. 2006 Mar;151(3):509-24. doi: 10.1007/s00705-005-0642-6. Epub 2005 Sep 30.
Translation initiation of hepatitis C virus RNA occurs via ribosome binding to an 'internal ribosome entry site (IRES)' located in the 5'untranslated region of the viral RNA. The principle interaction between the 40S ribosomal subunit and the HCV IRES has been shown to be largely factor independent, which is followed by the joining of the 60S ribosomal subunit to form functional 80S complex. However several additional cellular proteins have been reported to bind to HCV IRES and enhance the initiation of translation. In order to identify novel factors involved in the ribosome assembly during internal initiation of HCV RNA, northwestern screening of a HeLa cDNA expression library was performed, using HCV IRES RNA as probe. We demonstrate here, that human ribosomal protein L18a, a constituent of 60S subunit, interacts with HCV IRES RNA. This interaction was further confirmed by using a recombinant protein similar to L18a (sL18a), cloned from human blood. Interestingly, addition of increasing concentration of the purified recombinant sL18a protein, showed moderate stimulation of HCV IRES activity in the in vitro translation assay. These observations suggest that the human L18a might influence the HCV IRES mediated translation.
丙型肝炎病毒RNA的翻译起始是通过核糖体与位于病毒RNA 5'非翻译区的“内部核糖体进入位点(IRES)”结合来实现的。40S核糖体亚基与HCV IRES之间的主要相互作用已被证明在很大程度上不依赖于因子,随后60S核糖体亚基加入形成功能性80S复合物。然而,据报道有几种额外的细胞蛋白可与HCV IRES结合并增强翻译起始。为了鉴定在HCV RNA内部起始过程中参与核糖体组装的新因子,以HCV IRES RNA为探针,对HeLa cDNA表达文库进行了蛋白质印迹筛选。我们在此证明,60S亚基的组成成分人核糖体蛋白L18a与HCV IRES RNA相互作用。使用从人血液中克隆的类似于L18a的重组蛋白(sL18a)进一步证实了这种相互作用。有趣的是,在体外翻译试验中,添加浓度不断增加的纯化重组sL18a蛋白显示出对HCV IRES活性的适度刺激。这些观察结果表明,人L18a可能影响HCV IRES介导的翻译。