Biological significance of the second receptor binding site of Newcastle disease virus hemagglutinin-neuraminidase protein.
The paramyxovirus hemagglutinin-neuraminidase (HN) is a multifunctional protein responsible for attachment to receptors containing sialic acid, neuraminidase (NA) activity, and the promotion of membrane fusion, which is induced by the fusion protein. Analysis of the three-dimensional structure of Newcastle disease virus (NDV) HN protein revealed the presence of a large pocket, which mediates both receptor binding and NA activities. Recently, a second sialic acid binding site on HN was revealed by cocrystallization of the HN with a thiosialoside Neu5Ac-2-S-alpha(2,6)Gal1OMe, suggesting that NDV HN contains an additional sialic acid binding site. To evaluate the role of the second binding site on the life cycle of NDV, we rescued mutant viruses whose HNs were mutated at Arg516, a key residue that is involved in the second binding site. Loss of the second binding site on mutant HNs was confirmed by the hemagglutination inhibition test, which uses an inhibitor designed to block the NA active site. Characterization of the biological activities of HN showed that the mutation at Arg516 had no effect on NA activity. However, the fusion promotion activity of HN was substantially reduced by the mutation. Furthermore, the mutations at Arg516 slowed the growth rate of virus in tissue culture cells. These results suggest that the second binding site facilitates virus infection and growth by enhancing the fusion promotion activity of the HN.
Bousse
Tatiana L
TL
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.
Taylor
Garry
G
Krishnamurthy
Sateesh
S
Portner
Allen
A
Samal
Siba K
SK
Takimoto
Toru
T
D001665:Binding Sites; D002460:Cell Line; D015707:HN Protein / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D008561:Membrane Fusion; D009522:Newcastle disease virus / Q000737:chemistry* / Q000502:physiology; D011991:Receptors, Virus / Q000378:metabolism*
D015707:HN Protein; D011991:Receptors, Virus