title: Domain 3 of non-structural protein 5A from hepatitis C virus is natively unfolded.
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.
authors.affiliation: UGSF, UMR CNRS, IFR, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France. xavier.hanoulle@univ-lille1.fr
authors.forename: Xavier
authors.initials: X
authors.lastname: Hanoulle
authors.forename: Dries
authors.initials: D
authors.lastname: Verdegem
authors.forename: Aurélie
authors.initials: A
authors.lastname: Badillo
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Wieruszeski
authors.forename: François
authors.initials: F
authors.lastname: Penin
authors.forename: Guy
authors.initials: G
authors.lastname: Lippens
mesh_terms: D000595:Amino Acid Sequence; D016174:Hepacivirus / Q000378:metabolism*; D008969:Molecular Sequence Data; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase
chemical_list: D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase