Structural characterization of respiratory syncytial virus fusion inhibitor escape mutants: homology model of the F protein and a syncytium formation assay.
Respiratory syncytial virus (RSV) is a ubiquitous human pathogen and the leading cause of lower respiratory tract infections in infants. Infection of cells and subsequent formation of syncytia occur through membrane fusion mediated by the RSV fusion protein (RSV-F). A novel in vitro assay of recombinant RSV-F function has been devised and used to characterize a number of escape mutants for three known inhibitors of RSV-F that have been isolated. Homology modeling of the RSV-F structure has been carried out on the basis of a chimera derived from the crystal structures of the RSV-F core and a fragment from the orthologous fusion protein from Newcastle disease virus (NDV). The structure correlates well with the appearance of RSV-F in electron micrographs, and the residues identified as contributing to specific binding sites for several monoclonal antibodies are arranged in appropriate solvent-accessible clusters. The positions of the characterized resistance mutants in the model structure identify two promising regions for the design of fusion inhibitors.
Morton
Craig J
CJ
Biota Holdings Limited, 616 St Kilda Road, Melbourne, Victoria 3004, Australia. craig@medstv.unimelb.edu.au
Cameron
Rachel
R
Lawrence
Lynne J
LJ
Lin
Bo
B
Lowe
Melinda
M
Luttick
Angela
A
Mason
Anthony
A
McKimm-Breschkin
Jenny
J
Parker
Michael W
MW
Ryan
Jane
J
Smout
Michael
M
Sullivan
Jayne
J
Tucker
Simon P
SP
Young
Paul R
PR
D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001666:Binding Sites, Antibody; D002460:Cell Line; D015726:Giant Cells / Q000821:virology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D011487:Protein Conformation; D012136:Respiratory Syncytial Viruses / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D014760:Viral Fusion Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics* / Q000648:ultrastructure
D000998:Antiviral Agents; D014760:Viral Fusion Proteins