Translational Enhancement by the 3'-untranslated Region of Hepatitis C Virus RNA
Translational Enhancement by the 3'-untranslated Region of Hepatitis C Virus RNA
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DOI：
10.15369/sujms1989.15.99
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发表时间：
2003
期刊：
影响因子：
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通讯作者：
Kenichi Morikawa;Takayoshi Ito;H. Nozawa;Akio Miyokawa;Takeshi Saito;A. Katagiri;K. Mitamura
中科院分区：
文献类型：
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作者：
Kenichi Morikawa;Takayoshi Ito;H. Nozawa;Akio Miyokawa;Takeshi Saito;A. Katagiri;K. Mitamura
丙型肝炎病毒（HCV）RNA在5 '-非翻译区（UTR）中具有内部核糖体进入位点（IRES），在3'-UTR中具有X区。IRES和X区调节HCV翻译。HCV的X区通过与多聚嘧啶结合蛋白（PTB）的相互作用而增强其翻译。最近，几个细胞因子结合HCV RNA的两端被确定。然而，这些结合因子的功能仍不清楚。HCV RNA还含有一个长的poly（u/c）区，位于X区的上游。这个poly（U/C）区域在HCV翻译中的功能也不确定。在这项研究中，我们使用各种翻译系统分析了整个3 '-UTR对HCV翻译的作用。我们构建了基于T7的质粒，其含有5 '-UTR、作为报告基因的荧光素酶（Luc）基因和含有可变区、聚（U/ C）延伸和X区的整个3'-UTR（HCV 5 'L3'）。我们还制备了质粒，其中核心（HCV 5 'CL 3'）或核心至E2（HCV 5 'CE 1 E2 L3'）区域插入LUC基因之前。使用兔网织红细胞裂解物（RRL）的HCV 5 'CE 1 E2 L3'的体外翻译显示出整个3 '-UTR的25倍增强。我们还使用Huh 7、HeLa和HepG 2细胞在动物细胞培养系统中证明了3 '-UTR的这种翻译增强。总之，整个3 '-UTR可以增强兔网织红细胞裂解物和动物细胞培养系统中HCV RNA的翻译。
Hepatitis C virus (HCV) RNA has an internal ribosome entry site (IRES) in the 5'-untranslated region (UTR) and an X region in the 3'-UTR. The IRES and the X-region regulate HCV translation. HCV translation is enhanced by the X region through interaction with the polypyrimidine-tractbinding protein (PTB). Recently, several cellular factors that bind both ends of HCV RNA were identified. However, the function of these binding factors remains unclear. HCV RNA also contains a long poly (u/c) region just upstream of the X region. The function of this poly (U/C) region on HCV translation is also uncertain. In this study, we analyzed the role of the entire 3'-UTR on HCV translation using various translation systems. We constructed T7-based plasmids containing the 5'-UTR, the luciferase (Luc) gene as a reporter, and the entire 3'-UTR containing the variable region, poly (U/ C) stretch, and the X region (HCV 5'L3'). We also prepared plasmids in which the core (HCV 5'CL3') or the core-to-E2 (HCV 5'CE 1 E2L3') region was inserted preceding the LUC gene. In vitro translation of HCV 5'CE1E2L3' using rabbit reticulocyte lysates (RRL) showed a 25-fold enhancement by the entire 3'-UTR. We also demonstrated this translational enhancement by the 3'-UTR in animal cell culture systems using Huh7, HeLa, and HepG2 cells. In conclusion, the entire 3'-UTR can enhance translation of HCV RNA in both rabbit reticulocyte lysates and in animal cell culture systems.