Mutations in the Newcastle Disease Virus Hemagglutinin–Neuraminidase Protein That Interfere with Its Ability to Interact with the Homologous F Protein in the Promotion of Fusion

Deng, Ruitang; Wang, Zhiyu; Mahon, Paul J.; Marinello, Mark; Mirza, Anne M.; Iorio, Ronald M.

doi:10.1006/viro.1998.9501

Cited by 84 publications

(98 citation statements)

References 43 publications

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“…Our laboratory has shown that attachment-deficient NDV-AV HN, carrying substitutions for the catalytic aspartic acid, D198, is also fusion deficient and fails to interact with F at the cell surface in coimmunoprecipitation assays (4). This result is consistent with a mechanism for fusion in which receptor recognition by the globular domain of HN is the trigger for the HN-F interaction and fusion (17).…”

supporting

confidence: 60%

“…HN-expressing monolayers were incubated for 30 min at 4°C with a 2% suspension of erythrocytes in phosphate-buffered saline (PBS) supplemented with 1% CaCl 2 and MgCl 2 . After extensive washing, adsorbed erythrocytes were lysed in 50 mM NH 4 Cl and the lysate was clarified by centrifugation. HAd activity was quantitated by measuring the absorbance at 540 nm minus the background obtained with vector-expressing cells.…”

Section: Methodsmentioning

confidence: 99%

“…posttransfection, by a modification of a coimmunoprecipitation assay described previously (4). Because the amount of the F protein immunoprecipitated from the cell surface is greatly affected by the extent of fusion, we used a cleavage site mutant form of F for which cleavage activation and fusion are dependent on trypsin treatment.…”

Section: Vol 78 2004 Mutant Ndv Hn Interacts With F Despite Deficiementioning

confidence: 99%

“…A role for the HN stalk in fusion is also supported by the diminished fusogenic activity exhibited by HN proteins carrying point mutations in this region (4,25,27,30). The existence of an HN-F interaction is supported by the coimmunoprecipitation of the two proteins from the surface of both virusinfected and HN-F-cotransfected cells (4).…”

mentioning

confidence: 97%

“…This fusion activation requires a virus-specific interaction between the two proteins, the specificity of which is determined by a domain defined primarily by the stalk region of the HN spike, as shown by the analysis of chimeras composed of domains from heterologous HN proteins (5,32,33). A role for the HN stalk in fusion is also supported by the diminished fusogenic activity exhibited by HN proteins carrying point mutations in this region (4,25,27,30). The existence of an HN-F interaction is supported by the coimmunoprecipitation of the two proteins from the surface of both virusinfected and HN-F-cotransfected cells (4).…”

mentioning

confidence: 99%

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Mutated Form of the Newcastle Disease Virus Hemagglutinin-Neuraminidase Interacts with the Homologous Fusion Protein despite Deficiencies in both Receptor Recognition and Fusion Promotion

Lǐ

Quinlan

Mirza

et al. 2004

J Virol

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The Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein mediates attachment to cellular receptors. The fusion (F) protein promotes viral entry and spread. However, fusion is dependent on a virus-specific interaction between the two proteins that can be detected at the cell surface by a coimmunoprecipitation assay. A point mutation of I175E in the neuraminidase (NA) active site converts the HN of the Australia-Victoria isolate of the virus to a form that can interact with the F protein despite negligible receptor recognition and fusion-promoting activities. Thus, I175E-HN could represent a fusion intermediate in which HN and F are associated and primed for the promotion of fusion. Both the attachment and fusion-promoting activities of this mutant HN protein can be rescued either by NA activity contributed by another HN protein or by a set of four substitutions at the dimer interface. These substitutions were identified by the evaluation of chimeras composed of segments from HN proteins derived from two different NDV strains. These findings suggest that the I175E substitution converts HN to an F-interactive form, but it is one for which receptor binding is still required for fusion promotion. The data also indicate that the integrity of the HN dimer interface is critical to its receptor recognition activity.The Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) is a multifunctional, type II membrane glycoprotein. It exists at the surface of virions and virus-infected cells as a tetrameric spike structure, consisting of a long, membrane-proximal stalk region that supports a terminal globular domain. The latter region of the spike is responsible for HNЈs recognition of sialic acid-containing receptors as well as its neuraminidase (NA) activity, which cleaves sialic acid from both soluble and membrane-bound glycoconjugates (18). All of the antibody binding sites on the protein also reside in the globular domain (21).Several hundred strains of NDV have been isolated from all over the world. The X-ray crystallographic structures of the globular domain of the HN from one of these, the Kansas strain, as well as the protein complexed with sialic acid or an inhibitor of NA, have been solved (3). These structures suggest that the attachment and NA activities of HN are mediated by a single sialic acid binding site in two different conformations. Consistent with this, substitutions for several active site residues abolish both attachment and NA (3).Similarly, substitutions for residues in the NA active site of the HN from the Australia-Victoria (AV) isolate of NDV (NDV-AV) also result in the loss of detectable attachment and NA (11). These substitutions include I175E, D198R, K236R, Y526L, and E547Q. However, our laboratory has shown that the attachment activity of each of these receptor recognitiondeficient mutants can be partially rescued by providing NA activity (11). Clearly, this is inconsistent with the existence of a single site for both attachment and NA. Rather, it suggests that the defi...

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supporting

confidence: 60%

Section: Methodsmentioning

confidence: 99%

Section: Vol 78 2004 Mutant Ndv Hn Interacts With F Despite Deficiementioning

confidence: 99%

mentioning

confidence: 97%

mentioning

confidence: 99%

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Mutated Form of the Newcastle Disease Virus Hemagglutinin-Neuraminidase Interacts with the Homologous Fusion Protein despite Deficiencies in both Receptor Recognition and Fusion Promotion

Lǐ

Quinlan

Mirza

et al. 2004

J Virol

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Paramyxovirus Fusion (F) Protein: A Conformational Change on Cleavage Activation

et al. 2001

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The fusion (F) protein of the paramyxovirus SV5 promotes both virus-cell and cell-cell fusion. Recently, the atomic structure at 1.4 A of an extremely thermostable six-helix bundle core complex consisting of two heptad repeat regions of the F protein has been described (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S Jardetsky, Mol. Cell 3, 309-319, 1999). To analyze the conformations of the F protein at various stages of the membrane fusion process and to understand further the role of formation of the six-helix bundle core complex in promotion of membrane fusion, antibodies to peptides corresponding to regions of the F protein were obtained. Major changes in F protein antibody recognition were found after cleavage of the precursor protein F(0) to the fusogenically active disulfide-linked heterodimer, F(1) + F(2), and antibodies directed against the heptad repeat regions recognized only the uncleaved form. A monoclonal antibody directed against the F protein showed increased recognition at the cell surface of the cleaved form of the F protein as compared to uncleaved F protein, again indicating changes in conformation between the uncleaved and cleaved forms of the F protein. Anti-peptide antibodies specific for the heptad repeat regions were unable to precipitate a synthetic protein that consisted of the heptad repeat regions separated only by a small spacer, suggesting that the antibodies are unable to recognize their target regions when the heptad repeats are present in the six-helix bundle core complex. Taken together, these data indicate that the six-helix bundle core complex is not present in the precursor molecule F(0) and that significant conformational changes occur subsequent to cleavage of the F protein.

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Molecular Characterization of Menangle Virus, a Novel Paramyxovirus which Infects Pigs, Fruit Bats, and Humans

et al. 2001

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Menangle virus (MenV), isolated in August 1997 following an outbreak of reproductive disease in a piggery in New South Wales, is the second previously unclassified member of the family Paramyxoviridae to be identified in Australia since 1994. Similar to Hendra virus (HeV), MenV appears to be a virus of fruit bats (flying foxes) in the genus Pteropus. No serological cross-reactivity was detected between MenV and other known paramyxoviruses and to facilitate virus classification a cDNA subtraction method was used to obtain viral-specific cDNA from MenV-infected cells. Cloning and sequencing of the products enabled the entire sequences of the NP, P/V, M, F, and HN genes to be determined. Comparison of the nucleotide and deduced amino acid sequences for each gene with members of the family Paramyxoviridae, determination of the P gene mRNA editing strategy, and phylogenetic analyses confirmed that MenV is a new member of the genus Rubulavirus. However the deduced protein sequence of MenV HN exhibited only limited sequence homology when compared with attachment proteins of other paramyxoviruses. Key differences within the amino acid residues considered important determinants of neuraminidase activity suggest MenV HN is unlikely to possess the same degree of neuraminidase activity characteristic of other rubulavirus and respirovirus HN proteins.

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Mutations in the Newcastle Disease Virus Hemagglutinin–Neuraminidase Protein That Interfere with Its Ability to Interact with the Homologous F Protein in the Promotion of Fusion

Cited by 84 publications

References 43 publications

Mutated Form of the Newcastle Disease Virus Hemagglutinin-Neuraminidase Interacts with the Homologous Fusion Protein despite Deficiencies in both Receptor Recognition and Fusion Promotion

Mutated Form of the Newcastle Disease Virus Hemagglutinin-Neuraminidase Interacts with the Homologous Fusion Protein despite Deficiencies in both Receptor Recognition and Fusion Promotion

Paramyxovirus Fusion (F) Protein: A Conformational Change on Cleavage Activation

Molecular Characterization of Menangle Virus, a Novel Paramyxovirus which Infects Pigs, Fruit Bats, and Humans

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