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Monoclonal antibodies to the West Nile virus NS5 protein map to linear and conformational epitopes in the methyltransferase and polymerase domains
- Source :
- Journal of General Virology. 90:2912-2922
- Publication Year :
- 2009
- Publisher :
- Microbiology Society, 2009.
-
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.
- Subjects :
- Models, Molecular
RNA viruses
Protein Conformation
viruses
Viral Nonstructural Proteins
Binding, Competitive
Epitope
Mice
chemistry.chemical_compound
Kunjin virus
Virology
RNA polymerase
Animals
Humans
Binding site
Polymerase
Recombination, Genetic
Mice, Inbred BALB C
Binding Sites
biology
Linear epitope
Antibodies, Monoclonal
virus diseases
RNA
Methyltransferases
RNA-Dependent RNA Polymerase
biology.organism_classification
Molecular biology
chemistry
Mutation
biology.protein
Female
West Nile virus
Epitope Mapping
Conformational epitope
Subjects
Details
- ISSN :
- 14652099 and 00221317
- Volume :
- 90
- Database :
- OpenAIRE
- Journal :
- Journal of General Virology
- Accession number :
- edsair.doi.dedup.....97fc6b0148c41408bd3d7fa40df489a4