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Originally published In Press as doi:10.1074/jbc.M201722200 on March 7, 2002

J. Biol. Chem., Vol. 277, Issue 20, 17713-17721, May 17, 2002
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Alternate Translation Occurs within the Core Coding Region of the Hepatitis C Viral Genome*

Agoritsa VarakliotiDagger , Niki Vassilaki, Urania Georgopoulou, and Penelope Mavromara§

From the Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vassilisis Sofias Avenue, 115 21 Athens, Greece

The majority of hepatitis C virus (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 evidence supporting the expression of this ORF (designated core+1 ORF) via novel translation mechanisms. First, 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-luciferase) that exhibited ~54% luciferase activity relative to the positive control (core-luciferase). Second, antisera raised against two different synthetic core+1 peptides recognized the previously identified p16 (but not p21) core protein band expressed from HCV-1, indicating the presence of epitopes from the core+1 ORF within the p16 protein. Third, HCV-positive sera specifically recognized lysates of Escherichia coli cells 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 +1/-1 ribosomal frameshift 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.

* This work was supported by grants from the National Secretariat of Research and Technology. A preliminary report of this study describing the luciferase tagging experiments and the screening of human sera was presented at the Seventh International Meeting on Hepatitis C and Related Viruses, December 3-7, 2000, Gold Coast, Queensland, Australia.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Present address: Posttranscriptional Control Group, Dept. of Biomolecular Sciences, UMIST, P. O. Box 88, M60 1QD Manchester, UK.

§ To whom correspondence should be addressed. Tel.: 30-10-6478-877; Fax: 30-10-6478-877; E-mail: penelopm@hol.gr.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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