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Updated: Jun 25, 2026

A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Domain 3 of non-structural protein 5A from hepatitis C virus is natively unfolded
Xavier Hanoulle1, Dries Verdegem, Aurélie Badillo
1UGSF, UMR CNRS, IFR, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France. xavier.hanoulle@univ-lille1.fr
Abstract:
Hepatitis C virus (HCV) non-structural protein 5A (NS5A) is involved both in the viral replication and particle production. Its third domain (NS5A-D3), although not absolutely required for replication, is a key determinant for the production and assembly of novel HCV particles. As a prerequisite to elucidate the precise functions of this domain, we report here the first molecular characterization of purified recombinant HCV NS5A-D3. Sequence analysis indicates that NS5A-D3 is mostly unstructured but that short structural elements may exist at its N-terminus. Gel filtration chromatography, circular dichroism and finally NMR spectroscopy all point out the natively unfolded nature of purified recombinant NS5A-D3. This lack of stable folding is thought to be essential for primary interactions of NS5A-D3 domain with other viral or host proteins, which could stabilize some specific conformations conferring new functional features.
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