title: Unusual multiple recoding events leading to alternative forms of hepatitis C virus core protein from genotype 1b.
abstract: In addition to its involvement in the formation of the capsid shell of the virus particles, the core protein of hepatitis C virus (HCV) is believed to play an important role in the pathogenesis and/or establishment of persistent infection. We describe here alternative forms of genotype 1b HCV core protein identified after purification of various products of core protein segment 1-169 expressed in Escherichia coli and their analysis by proteolysis, mass spectrometry, and amino acid sequencing. These proteins all result from a +1 frameshift at codon 42 (a different position than that previously reported in genotype 1a) and, for some of them, from a rephasing in the normal open reading frame at the termination codon 144 in the +1 open reading frame. To test the relevance of these recoding events in a eukaryotic translational context, the nucleotide sequences surrounding the two shift sites were cloned in the three reading frames into expression vectors, allowing the production of a C-terminally fused green fluorescent protein, and expressed both in a reticulocyte lysate transcription/translation assay and in culture cells. Both recoding events were confirmed in these expression systems, strengthening the hypothesis that they might occur in HCV-infected cells. Moreover, sera from HCV-positive patients of genotype 1a or 1b were shown to react differently against synthetic peptides encoded in the +1 open reading frame. Together, these results indicate the occurrence of distinct recoding events in genotypes 1a and 1b, pointing out genotype-dependent specific features for F protein.
authors.affiliation: Laboratoire de Bioinformatique et RMN Structurales, aInstitut de Biologie et Chimie des Proteines, UMR5086 CNRS, Université Claude Bernard Lyon I, 7, Passage du Vercors, 69367 Lyon cedex 07, France.
authors.forename: Steeve
authors.initials: S
authors.lastname: Boulant
authors.forename: Michel
authors.initials: M
authors.lastname: Becchi
authors.forename: François
authors.initials: F
authors.lastname: Penin
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Lavergne
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003062:Codon; D004926:Escherichia coli / Q000378:metabolism; D016368:Frameshift Mutation; D005838:Genotype; D049452:Green Fluorescent Proteins; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000378:metabolism; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D014158:Transcription, Genetic; D014758:Viral Core Proteins / Q000737:chemistry* / Q000235:genetics*
chemical_list: D003062:Codon; D008164:Luminescent Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D049452:Green Fluorescent Proteins