Translation of human hepatitis C virus RNA in cultured cells is mediated by an internal ribosome-binding mechanism.
The human hepatitis C virus (HCV) contains a long 5' noncoding region (5' NCR). Computer-assisted and biochemical analyses suggest that there is a complex secondary structure in this region that is comparable to the secondary structures that are found in picornaviruses (E.A. Brown, H. Zhang, L.-H. Ping, and S.M. Lemon, Nucleic Acids Res. 20:5041-5045, 1992). Previous in vitro studies suggest that the HCV 5' NCR plays an important role during translation (K. Tsukiyama-Kohara, N. Iizuka, M. Kohara, and A. Nomoto, J. Virol. 66:1476-1483, 1992). Dicistronic and monocistronic expression vectors, in vitro translation, RNA transfections, and deletion mutagenesis studies were utilized to demonstrate unambiguously that the HCV 5' NCR is involved in translational control. Our data strongly support the conclusion that an internal ribosome entry site exists within the 5' noncoding sequences proximal to the initiator AUG. Furthermore, our results suggest that the HCV genome is translated in a cap-independent manner and that the sequences immediately upstream of the initiator AUG are essential for internal ribosome entry site function during translation.
Department of Microbiology and Immunology, University of Colorado Medical School, Denver 80262.
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Wang
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Sarnow
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Siddiqui
D001483:Base Sequence; D003062:Codon; D004252:DNA Mutational Analysis; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010442:Peptide Chain Initiation, Translational; D010957:Plasmids / Q000235:genetics; D017955:Poliovirus / Q000254:growth & development; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism*; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D015678:Viral Structural Proteins / Q000235:genetics
D003062:Codon; D012367:RNA, Viral; D015678:Viral Structural Proteins