Stem-loop structures II-IV of the 5' untranslated sequences are required for the expression of the full-length hepatitis C virus genome
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 g...
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| Veröffentlicht in: | Archives of virology 2003-03, Vol.148 (3), p.449-467 |
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| creator | QI, Z. T KALKERI, G HANIBLE, J PRABHU, R BASTIAN, F GARRY, R. F DASH, S |
| description | 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. |
| doi_str_mv | 10.1007/s00705-002-0933-0 |
| format | Article |
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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. 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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. 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T</au><au>KALKERI, G</au><au>HANIBLE, J</au><au>PRABHU, R</au><au>BASTIAN, F</au><au>GARRY, R. F</au><au>DASH, S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Stem-loop structures II-IV of the 5' untranslated sequences are required for the expression of the full-length hepatitis C virus genome</atitle><jtitle>Archives of virology</jtitle><addtitle>Arch Virol</addtitle><date>2003-03-01</date><risdate>2003</risdate><volume>148</volume><issue>3</issue><spage>449</spage><epage>467</epage><pages>449-467</pages><issn>0304-8608</issn><eissn>1432-8798</eissn><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. 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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.</abstract><cop>Wien</cop><cop>New York, NY</cop><pub>Springer</pub><pmid>12607098</pmid><doi>10.1007/s00705-002-0933-0</doi><tpages>19</tpages></addata></record> |
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| subjects | 3' Untranslated Regions - chemistry 3' Untranslated Regions - genetics 5' Untranslated Regions - chemistry 5' Untranslated Regions - genetics Amino acids Base Sequence Biological and medical sciences Cell Line Cloning Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Regulation, Viral Genetics Genome, Viral Genomes Green Fluorescent Proteins Health sciences Hepacivirus - genetics Hepacivirus - metabolism Hepatitis C Humans Luminescent Proteins - genetics Luminescent Proteins - metabolism Microbiology Molecular Sequence Data Mutation Protein Biosynthesis Protein expression Proteins Recombinant Fusion Proteins - metabolism Ribonucleic acid RNA RNA polymerase RNA, Viral - metabolism Viral Proteins - genetics Viral Proteins - metabolism Virology Viruses |
| title | 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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