Identification of amino acid residues important to the neuraminidase activity of the HN glycoprotein of Newcastle disease virus.
Monoclonal antibodies (MAbs) to three overlapping antigenic sites (designated 12, 2, and 23) on the hemagglutinin-neuraminidase glycoprotein (HN) of Newcastle disease virus (NDV) were previously shown to inhibit neuraminidase activity (NA) on neuraminlactose (R. M. Iorio and M. A. Bratt, 1984a, J. Immunol. 133, 2215-2219; R. M. Iorio et al., 1989, Virus Res. 13, 245-262). However, a competitive inhibitor of NA blocks the binding of only MAbs to site 23, suggesting that the domain they recognize may be closely related to the NA site. Antigenic variants selected with site 23 MAbs have single amino acid substitutions at HN residues 192, 193, or 200. Virions of variants, which have a substitution at residue 193 or 200, have alterations in NA which are not attributable to a commensurate change in HN content. A revertant of a temperature-sensitive mutant, which has markedly diminished NA relative to the wild type, has an amino acid substitution at residue 175. A second step revertant having partially restored NA has an additional substitution at residue 192 identical to that in one of the site 23 variants, which, in turn, also makes the revertant resistant to neutralization by site 23 MAbs. Thus, an amino acid substitution at residue 175, 193, or 200 of the HN of NDV can have marked effects on the NA of the protein. The amino acids in the region around residue 175 are highly conserved between the HNs of NDV and other paramyxoviruses, suggesting that this domain is important to the integrity of the NA site in this group of viruses.
Iorio
R M
RM
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester 01655.
Syddall
R J
RJ
Glickman
R L
RL
Riel
A M
AM
Sheehan
J P
JP
Bratt
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D001483:Base Sequence; D001667:Binding, Competitive; D002642:Chick Embryo; D000939:Epitopes / Q000276:immunology; D015707:HN Protein / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000031:analogs & derivatives*; D009439:Neuraminidase / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D009522:Newcastle disease virus / Q000201:enzymology* / Q000276:immunology; D012794:Sialic Acids / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D015707:HN Protein; D012794:Sialic Acids; C036137:2-deoxy-2,3-dehydro-N-acetylneuraminic acid; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid