Domain 3 of NS5A protein from the hepatitis C virus has intrinsic α-helical propensity and is a substrate of cyclophilin A

• Verfasst von: Verdegem, Dries [VerfasserIn]
• Verfasst von: Badillo, Aurélie [VerfasserIn]
• Verfasst von: Wieruszeski, Jean-Michel [VerfasserIn]
• Verfasst von: Landrieu, Isabelle [VerfasserIn]
• Verfasst von: Leroy, Arnaud [VerfasserIn]
• Verfasst von: Bartenschlager, Ralf [VerfasserIn]
• Verfasst von: Penin, François [VerfasserIn]
• Verfasst von: Lippens, Guy [VerfasserIn]
• Verfasst von: Hanoulle, Xavier [VerfasserIn]
• Titel: Domain 3 of NS5A protein from the hepatitis C virus has intrinsic α-helical propensity and is a substrate of cyclophilin A
• Verf.angabe: Dries Verdegem, Aurélie Badillo, Jean-Michel Wieruszeski, Isabelle Landrieu, Arnaud Leroy, Ralf Bartenschlager, François Penin, Guy Lippens, and Xavier Hanoulle
• E-Jahr: 2011
• Jahr: 13 April 2011
• Umfang: 14 S.
• Fussnoten: Gesehen am 07.12.2022
• Titel Quelle: Enthalten in: The journal of biological chemistry
• Ort Quelle: Bethesda, Md. : ASBMB Publications, 1905
• Jahr Quelle: 2011
• Band/Heft Quelle: 286(2011), 23, Seite 20441-20454
• ISSN Quelle: 1083-351X
• DOI: doi:10.1074/jbc.M110.182436
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: HCV
• Sach-SW: NMR
• Sach-SW: Prolyl Isomerase
• Sach-SW: Protein Domains
• Sach-SW: Protein Structure
• Sach-SW: Protein-Protein Interactions
• K10plus-PPN: 1826466568

Abstract

Nonstructural protein 5A (NS5A) is essential for hepatitis C virus (HCV) replication and constitutes an attractive target for antiviral drug development. Although structural data for its in-plane membrane anchor and domain D1 are available, the structure of domains 2 (D2) and 3 (D3) remain poorly defined. We report here a comparative molecular characterization of the NS5A-D3 domains of the HCV JFH-1 (genotype 2a) and Con1 (genotype 1b) strains. Combining gel filtration, CD, and NMR spectroscopy analyses, we show that NS5A-D3 is natively unfolded. However, NS5A-D3 domains from both JFH-1 and Con1 strains exhibit a propensity to partially fold into an α-helix. NMR analysis identifies two putative α-helices, for which a molecular model could be obtained. The amphipathic nature of the first helix and its conservation in all genotypes suggest that it might correspond to a molecular recognition element and, as such, promote the interaction with relevant biological partner(s). Because mutations conferring resistance to cyclophilin inhibitors have been mapped into NS5A-D3, we also investigated the functional interaction between NS5A-D3 and cyclophilin A (CypA). CypA indeed interacts with NS5A-D3, and this interaction is completely abolished by cyclosporin A. NMR heteronuclear exchange experiments demonstrate that CypA has in vitro peptidyl-prolyl cis/trans-isomerase activity toward some, but not all, of the peptidyl-prolyl bonds in NS5A-D3. These studies lead to novel insights into the structural features of NS5A-D3 and its relationships with CypA.