A single amino acid substitution in the haemagglutinin-neuraminidase protein of Newcastle disease virus results in increased fusion promotion and decreased neuraminidase activities without changes in virus pathotype.
Attachment of Newcastle disease virus (NDV) to the host cell is mediated by the haemagglutinin-neuraminidase (HN), a multifunctional protein that has receptor recognition, neuraminidase (NA) and fusion promotion activities. The process that connects receptor binding and fusion triggering is poorly understood and amino acid residues important for the functions of the protein remain to be fully determined. During the process of generating an infectious clone of the Anhinga strain of NDV, we were able to rescue a NDV with highly increased fusogenic activity in vitro and decreased haemagglutinating activity, as compared with the wild-type parental strain. Sequencing of this recombinant virus showed a single mutation at amino acid position 192 of the HN protein (Ile→Met). In the present study, we characterized that single amino acid substitution (I192M) in three strains of NDV by assessing the NA activity and fusogenic potential of the mutated versus wild-type proteins in cell cultures. The original recombinant NDV harbouring the mutation in the HN gene was also used to characterize the phenotype of the virus in cell cultures, embryonated chicken eggs and day-old chickens. Mutation I192M results in low NA activity and highly increased cell fusion in vitro, without changes in the viral pathotype of recombinant viruses harbouring the mutation in vivo. The results obtained suggest that multiple regions of the HN-protein globular head are important for fusion promotion, and that wild-type levels of NA activity are not absolutely required for viral infection.
Southeast Poultry Research Laboratory, Agricultural Research Service, United States Department of Agriculture, 934 College Station Road, Athens, GA 30605, USA.
Carlos
C
Estevez
Daniel J
DJ
King
Ming
M
Luo
Qingzhong
Q
Yu
D019943:Amino Acid Substitution / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D002522:Chlorocebus aethiops; D004252:DNA Mutational Analysis; D015707:HN Protein / Q000235:genetics*; D006801:Humans; D020125:Mutation, Missense; D009521:Newcastle Disease / Q000401:mortality / Q000473:pathology / Q000821:virology; D009522:Newcastle disease virus / Q000472:pathogenicity* / Q000502:physiology*; D017422:Sequence Analysis, DNA; D016019:Survival Analysis; D014774:Virulence; D053586:Virus Internalization
D015707:HN Protein