Stem-loop structures II-IV of the 5? untranslated sequences are required for the expression of the full-length hepatitis C virus genome

Autor: Gururaj Kalkeri, Ramesh Prabhu, Robert F. Garry, Z. T. Qi, J. Hanible, Frank Bastian, Srikanta Dash
Rok vydání: 2003
Předmět:
Zdroj: Archives of Virology. 148:449-467
ISSN: 1432-8798
0304-8608
DOI: 10.1007/s00705-002-0933-0
Popis: The 5' and 3' untranslated regions (UTR) of the hepatitis C virus (HCV) genome contain stem-loop structures, which are important in viral gene expression and replication. In this study, the functional roles of the predicted stem-loop structures of HCV 5' UTR and 3' UTR in viral gene expression were examined using a chimeric clone of full-length HCV genomic cDNA clone and the gene for green fluorescent protein (GFP). High level expression of the HCV-GFP chimera in Huh-7 cells was accomplished by using a replication defective adenovirus that expresses T7 RNA polymerase and transcription plasmid containing full-length HCV-GFP chimera under the control of a T7 promoter. The HCV-GFP clone, with deletion of stem-loop I, expressed proteins in transfected Huh-7 cells at comparable levels to the wild type HCV clone. Other mutations of the 5' UTR, which either deleted or altered the base pairing of stem-loops II to IV, completely abolished the expression of HCV-GFP chimera. In contrast, deletion of 3' UTR sequences had no effect on HCV protein expression. These findings suggest that the stem-loop structures II to IV of HCV 5' UTR are necessary for protein expression, but that stem loop I is dispensable for protein translation. The stem-loop structures of 3' UTR of HCV genome appear to have no direct role in viral gene expression.
Databáze: OpenAIRE
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