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Archives of Virology

, Volume 148, Issue 3, pp 449–467 | Cite as

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

  • Z. T. Qi
  • G. Kalkeri
  • J. Hanible
  • R. Prabhu
  • F. Bastian
  • R. F. Garry
  • S. Dash

Summary.

 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.

Keywords

Viral Gene Expression Untranslated Sequence Genomic cDNA Replication Defective Adenovirus Chimeric Clone 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag/Wien 2002

Authors and Affiliations

  • Z. T. Qi
    • 1
  • G. Kalkeri
    • 1
  • J. Hanible
    • 1
  • R. Prabhu
    • 1
  • F. Bastian
    • 1
  • R. F. Garry
    • 2
  • S. Dash
    • 1
  1. 1.Department of Pathology and Laboratory Medicine, Tulane University Health Sciences Center, New Orleans, U.S.A.US
  2. 2.Department of Microbiology and Immunology, Tulane University Health Sciences Center, New Orleans, U.S.A.US

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