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

Stem-loop structures II–IV of the 5′ untranslated sequences are required for the expression... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

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

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Publisher
Springer Journals
Copyright
Copyright © 2002 by Springer-Verlag/Wien
Subject
Legacy
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-002-0933-0
Publisher site
See Article on Publisher Site

Abstract

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.

Journal

Archives of VirologySpringer Journals

Published: Feb 1, 2003

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