Title |
Monoclonal antibodies to the West Nile virus NS5 protein map to linear and conformational epitopes in the methyltransferase and polymerase domains
|
---|---|
Published in |
Journal of General Virology, August 2009
|
DOI | 10.1099/vir.0.013805-0 |
Pubmed ID | |
Authors |
Roy A Hall, Si En Tan, Barbara Selisko, Rachael Slade, Jody Hobson-Peters, Bruno Canard, Megan Hughes, Jason Y Leung, Ezequiel Balmori-Melian, Sonja Hall-Mendelin, Kim B Pham, David C Clark, Natalie A Prow, Alexander A Khromykh |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Netherlands | 1 | 3% |
France | 1 | 3% |
Germany | 1 | 3% |
Unknown | 32 | 91% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 9 | 26% |
Researcher | 8 | 23% |
Student > Master | 4 | 11% |
Student > Bachelor | 3 | 9% |
Student > Postgraduate | 2 | 6% |
Other | 5 | 14% |
Unknown | 4 | 11% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 12 | 34% |
Medicine and Dentistry | 5 | 14% |
Biochemistry, Genetics and Molecular Biology | 5 | 14% |
Veterinary Science and Veterinary Medicine | 3 | 9% |
Immunology and Microbiology | 3 | 9% |
Other | 3 | 9% |
Unknown | 4 | 11% |