title: Loss of N-linked glycosylation from the hemagglutinin-neuraminidase protein alters virulence of Newcastle disease virus.
abstract: The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) is an important determinant of its virulence. We investigated the role of each of the four functional N-linked glycosylation sites (G1 to G4) of the HN glycoprotein of NDV on its pathogenicity. The N-linked glycosylation sites G1 to G4 at residues 119, 341, 433, and 481, respectively, of a moderately pathogenic NDV strain Beaudette C (BC) were eliminated individually by site-directed mutagenesis on a full-length cDNA clone of BC. A double mutant (G12) was also created by eliminating the first and second glycosylation sites at residues 119 and 341, respectively. Infectious virus was recovered from each of the cDNA clones of the HN glycoprotein mutants, employing a reverse genetics technique. There was a greater delay in the replication of G4 and G12 mutant viruses than in the parental virus. Loss of glycosylation does not affect the receptor recognition by HN glycoprotein of NDV. The neuraminidase activity of G4 and G12 mutant viruses and the fusogenicity of the G4 mutant virus were significantly lower than those of the parental virus. The fusogenicity of the double mutant virus (G12) was significantly higher than that of the parental virus. Cell surface expression of the G4 virus HN was significantly lower than that of the parental virus. The antigenic reactivities of the mutants to a panel of monoclonal antibodies against the HN protein indicated that removal of glycosylation from the HN protein increased (G1, G3, and G12) or decreased (G2 and G4) the formation of antigenic sites, depending on their location. In standard tests to assess virulence in chickens, all of the glycosylation mutants were less virulent than the parental BC virus, but the G4 and G12 mutants were the least virulent.
authors.affiliation: Virginia-Maryland Regional College of Veterinary Medicine, University of Maryland, 8075 Greenmead Drive, College Park, MD 20742, USA.
authors.forename: Aruna
authors.initials: A
authors.lastname: Panda
authors.forename: Subbiah
authors.initials: S
authors.lastname: Elankumaran
authors.forename: Sateesh
authors.initials: S
authors.lastname: Krishnamurthy
authors.forename: Zhuhui
authors.initials: Z
authors.lastname: Huang
authors.forename: Siba K
authors.initials: SK
authors.lastname: Samal
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D006031:Glycosylation; D015707:HN Protein / Q000737:chemistry* / Q000502:physiology; D009154:Mutation; D009522:Newcastle disease virus / Q000737:chemistry / Q000472:pathogenicity*; D011233:Precipitin Tests; D014774:Virulence
chemical_list: D000911:Antibodies, Monoclonal; D015707:HN Protein