Monoclonal antibodies to the West Nile virus NS5 protein map to linear and conformational epitopes in the methyltransferase and polymerase domains

Hall, Roy A., Tan, Si En, Selisko, Barbara, Slade, Rachael, Hobson-Peters, Jody, Canard, Bruno, Hughes, Megan, Leung, Jason Y., Balmori-Melian, Ezequiel, Hall-Mendelin, Sonja, Pham, Kim B., Clark, David C., Prow, Natalie A. and Khromykh, Alexander A. (2009) Monoclonal antibodies to the West Nile virus NS5 protein map to linear and conformational epitopes in the methyltransferase and polymerase domains. Journal of General Virology, 90 12: 2912-2922. doi:10.1099/vir.0.013805-0


Author Hall, Roy A.
Tan, Si En
Selisko, Barbara
Slade, Rachael
Hobson-Peters, Jody
Canard, Bruno
Hughes, Megan
Leung, Jason Y.
Balmori-Melian, Ezequiel
Hall-Mendelin, Sonja
Pham, Kim B.
Clark, David C.
Prow, Natalie A.
Khromykh, Alexander A.
Title Monoclonal antibodies to the West Nile virus NS5 protein map to linear and conformational epitopes in the methyltransferase and polymerase domains
Journal name Journal of General Virology   Check publisher's open access policy
ISSN 0022-1317
1465-2099
Publication date 2009-12
Sub-type Article (original research)
DOI 10.1099/vir.0.013805-0
Volume 90
Issue 12
Start page 2912
End page 2922
Total pages 11
Editor R. M. Elliot
Place of publication Reading, U.K.
Publisher Society for General Microbiology
Collection year 2010
Language eng
Subject C1
060502 Infectious Agents
060506 Virology
920109 Infectious Diseases
Formatted abstract
The West Nile virus (WNV) NS5 protein contains a methyltransferase (MTase) domain involved in RNA capping and an RNA-dependent RNA polymerase (RdRp) domain essential for virus replication. Crystal structures of individual WNV MTase and RdRp domains have been solved; however, the structure of full-length NS5 has not been determined. To gain more insight into the structure of NS5 and interactions between the MTase and RdRp domains, we generated a panel of seven monoclonal antibodies (mAbs) to the NS5 protein of WNV (Kunjin strain) and mapped their binding sites using a series of truncated NS5 proteins and synthetic peptides. Binding sites of four mAbs (5D4, 4B6, 5C11 and 6A10) were mapped to residues 354–389 in the fingers subdomain of the RdRp. This is consistent with the ability of these mAbs to inhibit RdRp activity in vitro and suggests that this region represents a potential target for RdRp inhibitors. Using a series of synthetic peptides, we also identified a linear epitope (bound by mAb 5H1) that mapped to a 13 aa stretch surrounding residues 47 and 49 in the MTase domain, a region predicted to interact with the palm subdomain of the RdRp. The failure of one mAb (7G6) to bind both N- and C-terminally truncated NS5 recombinants indicates that the antibody recognizes a conformational epitope that requires the presence of residues in both the MTase and RdRp domains. These data support a structural model of the full-length NS5 molecule that predicts a physical interaction between the MTase and the RdRp domains.
© 2009 SGM
Keyword West Nile virus
Valley encephalitis-virus
RNA-synthesis
Kunjin virus
Nonstructural protein-5
Escherichia-coli
In-vitro
Dengue
Glycoproteins
Infection
Envelope
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
School of Chemistry and Molecular Biosciences
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 18 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 18 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Sun, 13 Dec 2009, 00:03:34 EST