Mutations in the ectodomain of newcastle disease virus fusion protein confer a hemagglutinin-neuraminidase-independent phenotype.
The entry of enveloped viruses into host cells is preceded by membrane fusion, which in paramyxoviruses is triggered by the fusion (F) protein. Refolding of the F protein from a metastable conformation to a highly stable postfusion form is critical for the promotion of fusion, although the mechanism is still not well understood. Here we examined the effects of mutations of individual residues of the F protein of Newcastle disease virus, located at critical regions of the protein, such as the C terminus of the N-terminal heptad repeat (HRA) and the N terminus of the C-terminal heptad repeat (HRB). Seven of the mutants were expressed at the cell surface, showing differences in antibody reactivity in comparison with the F wild type. The N211A, L461A, I463A, and I463F mutants showed a hyperfusogenic phenotype both in syncytium and in dye transfer assays. The four mutants promoted fusion more efficiently at lower temperatures than the wild type did, meaning they probably had lower energy requirements for activation. Moreover, the N211A, I463A, and I463F mutants exhibited hemagglutinin-neuraminidase (HN)-independent activity when influenza virus hemagglutinin (HA) was coexpressed as an attachment protein. The data are discussed in terms of alterations of the refolding pathway and/or the stability of the prefusion and fusion conformations.
Ayllón
Juan
J
Departamento de Bioquímica y Biología Molecular, Universidad de Salamanca, Edificio Departamental Lab. 112, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain.
Villar
Enrique
E
Muñoz-Barroso
Isabel
I
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D015726:Giant Cells / Q000378:metabolism; D015707:HN Protein / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008561:Membrane Fusion; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009522:Newcastle disease virus / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D011487:Protein Conformation; D014162:Transfection; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000378:metabolism
D015707:HN Protein; D014760:Viral Fusion Proteins