Crystal structure of the RNA polymerase domain of the West Nile virus non-structural protein 5.
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.
Malet
Hélène
H
Architecture et Fonction des Macromolécules Biologiques, CNRS, and Universités d'Aix-Marseille I et II, UMR 6098, ESIL Case 925, 13288 Marseille, France.
Egloff
Marie-Pierre
MP
Selisko
Barbara
B
Butcher
Rebecca E
RE
Wright
Peter J
PJ
Roberts
Michael
M
Gruez
Arnaud
A
Sulzenbacher
Gerlind
G
Vonrhein
Clemens
C
Bricogne
Gérard
G
Mackenzie
Jason M
JM
Khromykh
Alexander A
AA
Davidson
Andrew D
AD
Canard
Bruno
B
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
D004791:Enzyme Inhibitors; C096041:NS5 protein, flavivirus; D017361:Viral Nonstructural Proteins; D012359:tRNA Methyltransferases; D012324:RNA-Dependent RNA Polymerase