In vitro selection of the 3鈥?untranslated regions of the human liver mRNA that bind to the HCV nonstructural protein 5B

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• 作者：Kazuhito Yuhashi ; Shin Ohnishi ; Tatsuhiko Kodama ; Kazuhiko Koike ; Hiroshi Kanamori
• 关键词：HCV ; NS5B ; LGALS1 (GAL-1) ; mRNA 3&prime ; -UTR ; RPS4X
• 刊名：Virology
• 出版年：February, 2014
• 年：2014
• 卷：450-451
• 期：Complete
• 页码：13-23
• 全文大小：1218 K

文摘

Hepatitis C virus (HCV) nonstructural protein 5B (NS5B) has RNA-dependent RNA polymerase (RdRp) activity. Because NS5B recognizes various RNA motifs besides the HCV genome, NS5B has the potential of interacting with host RNA molecules. In this study, an RNA pool enriched with the 3鈥?UTR sequences was generated and mRNA molecules with high affinity binding to NS5B were selected by iterative selection. Among the high binding mRNA 3鈥?UTR segments, we analyzed the housekeeping ribosomal protein S4, X-linked [RPS4X] mRNA 3鈥?UTR and the 3鈥?UTR of galectin-1 (GAL-1) mRNA, which is known to be one of the genes upregulated in HCV-infected liver cells and to have a wide spectrum of biological properties. By means of IP-RT-PCR, it was demonstrated that both of the mRNA molecules bind to NS5B in the cytoplasm. Interestingly, GAL-1 and RPS4X mRNA can serve as templates for NS5B RdRp, suggesting these RNA molecules are regulated in vivo by NS5B.