title: Crystal structure of the RNA polymerase domain of the West Nile virus non-structural protein 5.
abstract: Viruses of the family Flaviviridae are important human and animal pathogens. Among them, the Flaviviruses dengue (DENV) and West Nile (WNV) cause regular outbreaks with fatal outcomes. The RNA-dependent RNA polymerase (RdRp) activity of the non-structural protein 5 (NS5) is a key activity for viral RNA replication. In this study, crystal structures of enzymatically active and inactive WNV RdRp domains were determined at 3.0- and 2.35-A resolution, respectively. The determined structures were shown to be mostly similar to the RdRps of the Flaviviridae members hepatitis C and bovine viral diarrhea virus, although with unique elements characteristic for the WNV RdRp. Using a reverse genetic system, residues involved in putative interactions between the RNA-cap methyltransferase (MTase) and the RdRp domain of Flavivirus NS5 were identified. This allowed us to propose a model for the structure of the full-length WNV NS5 by in silico docking of the WNV MTase domain (modeled from our previously determined structure of the DENV MTase domain) onto the RdRp domain. The Flavivirus RdRp domain structure determined here should facilitate both the design of anti-Flavivirus drugs and structure-function studies of the Flavivirus replication complex in which the multifunctional NS5 protein plays a central role.
authors.affiliation: Architecture et Fonction des Macromolécules Biologiques, CNRS, and Universités d'Aix-Marseille I et II, UMR 6098, ESIL Case 925, 13288 Marseille, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Malet
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Egloff
authors.forename: Barbara
authors.initials: B
authors.lastname: Selisko
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Butcher
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Wright
authors.forename: Michael
authors.initials: M
authors.lastname: Roberts
authors.forename: Arnaud
authors.initials: A
authors.lastname: Gruez
authors.forename: Gerlind
authors.initials: G
authors.lastname: Sulzenbacher
authors.forename: Clemens
authors.initials: C
authors.lastname: Vonrhein
authors.forename: Gérard
authors.initials: G
authors.lastname: Bricogne
authors.forename: Jason M
authors.initials: JM
authors.lastname: Mackenzie
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Khromykh
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Davidson
authors.forename: Bruno
authors.initials: B
authors.lastname: Canard
mesh_terms: D000818:Animals; D018360:Crystallography, X-Ray; D001908:Diarrhea Viruses, Bovine Viral / Q000737:chemistry / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000627:therapeutic use; D016174:Hepacivirus / Q000737:chemistry / Q000201:enzymology; D006801:Humans; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D040681:Structural Homology, Protein; D013329:Structure-Activity Relationship; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D014901:West Nile Fever / Q000188:drug therapy / Q000201:enzymology; D014902:West Nile virus / Q000201:enzymology*; D012359:tRNA Methyltransferases / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; C096041:NS5 protein, flavivirus; D017361:Viral Nonstructural Proteins; D012359:tRNA Methyltransferases; D012324:RNA-Dependent RNA Polymerase