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Papers In Press, published online ahead of print March 7, 2002
Molecular Virology Laboratory, Hellenic Pasteur Institute, Athens 157 72
Corresponding Author: penelopm{at}hol.gr
The majority of HCV isolates contain an open reading frame (ORF) overlapping with the core coding sequences in the +1 frame which was assumed to be untranslated. We present here evidence supporting the expression of this ORF, designated as core+1 ORF, via novel translation mechanisms. Firstly, fusion of the luciferase gene with the HCV-1 core+1 ORF followed by in vitro translation resulted in the synthesis of a chimeric protein (core+1-luc), which exhibited luciferase activity of about 54% relative to the positive control (core-luc). Secondly, antisera raised against two different synthetic core+1 peptides recognised the previously identified P16, but not the P21, core protein band expressed from the HCV-1, indicating the presence of epitopes from the core+1 ORF within the P16 protein. Thirdly, HCV positive sera specifically recognised lysates of Escherichia coli expressing recombinant core+1 protein, suggesting the presence of anti-core+1 antibodies in HCV-infected patients. Finally, luciferase tagging experiments designed to assess for -1 frameshifting combined with site-directed mutagenesis experiments supported the presence of both +1/-1 ribosomal frameshifting translation mechanisms within the core coding region. In conclusion our data provide evidence for novel translation mechanisms within the core-coding region and demonstrate the expression of the core+1 ORF, at least for some HCV isolates.
J. Biol. Chem, 10.1074/jbc.M201722200
Submitted on February 20, 2002
Revised on February 28, 2002
Accepted on March 7, 2002
Alternate translation occurs within the core coding region of the hepatitis C viral genome
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