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Prevalence of intrinsic disorder in the hepatitis C virus ARFP/Core+1/S protein
Author(s) -
Boumlic Anissa,
Nominé Yves,
Charbonnier Sebastian,
Dalagiorgou Georgia,
Vassilaki Niki,
Kieffer Bruno,
Travé Gilles,
Mavromara Penelope,
Orfanoudakis Georges
Publication year - 2010
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2009.07527.x
Subject(s) - core protein , recombinant dna , open reading frame , virology , genome , core (optical fiber) , size exclusion chromatography , escherichia coli , biology , chemistry , computational biology , virus , genetics , microbiology and biotechnology , peptide sequence , gene , biochemistry , enzyme , physics , optics
The hepatitis C virus (HCV) Core+1/S polypeptide, also known as alternative reading frame protein (ARFP)/S, is an ARFP expressed from the Core coding region of the viral genome. Core+1/S is expressed as a result of internal initiation at AUG codons (85–87) located downstream of the polyprotein initiator codon, and corresponds to the C‐terminal part of most ARFPs. Core+1/S is a highly basic polypeptide, and its function still remains unclear. In this work, untagged recombinant Core+1/S was expressed and purified from Escherichia coli in native conditions, and was shown to react with sera of HCV‐positive patients. We subsequently undertook the biochemical and biophysical characterization of Core+1/S. The conformation and oligomeric state of Core+1/S were investigated using size exclusion chromatography, dynamic light scattering, fluorescence, CD, and NMR. Consistent with sequence‐based disorder predictions, Core+1/S lacks significant secondary structure in vitro , which might be relevant for the recognition of diverse molecular partners and/or for the assembly of Core+1/S. This study is the first reported structural characterization of an HCV ARFP/Core+1 protein, and provides evidence that ARFP/Core+1/S is highly disordered under native conditions, with a tendency for self‐association.