Abstract
The N1 and N9 neuraminidase (NA) subtypes of influenza A viruses exhibit significant hemadsorption activity that localizes to a site distinct from that of the enzymatic active site. To determine the conservation of hemadsorption activity among different NAs, we have examined most of the NA subtypes from avian, swine, equine, and human virus isolates. All subtypes of avian virus NAs examined and one equine virus N8 NA possessed high levels of hemadsorption activity. A swine virus N1 NA exhibited only weak hemadsorption activity, while in human virus N1 and N2 NAs, the activity was detected at a much lower level than in avian virus NAs. NAs which possessed hemadsorption activity for chicken erythrocytes (RBCs) were similarly able to adsorb human RBCs. However, none of the hemadsorption-positive NAs could bind equine, swine, or bovine RBCs, suggesting that RBCs from these species lack molecules, recognized by the NA hemadsorption site, present on human and chicken RBCs. Mutagenesis of the putative hemadsorption site of A/duck/Hong Kong/7/75 N2 NA abolished the high level of hemadsorption activity exhibited by the wild-type protein but also resulted in a 50% reduction of the NA enzymatic activity. A transfectant virus, generated by reverse genetics, containing this mutated NA replicated 10-fold less efficiently in chicken embryo fibroblast cultures than did a transfectant virus expressing the wild-type NA. However, both viruses replicated equally well in Peking ducks. Although conservation of NA hemadsorption activity among avian virus NAs suggests the maintenance of a required function of NA, loss of the activity does not preclude the replication of the virus in an avian host.
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Selected References
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- Air G. M., Laver W. G. Red cells bound to influenza virus N9 neuraminidase are not released by the N9 neuraminidase activity. Virology. 1995 Aug 1;211(1):278–284. doi: 10.1006/viro.1995.1401. [DOI] [PubMed] [Google Scholar]
- EYLAR E. H., MADOFF M. A., BRODY O. V., ONCLEY J. L. The contribution of sialic acid to the surface charge of the erythrocyte. J Biol Chem. 1962 Jun;237:1992–2000. [PubMed] [Google Scholar]
- Enami M., Luytjes W., Krystal M., Palese P. Introduction of site-specific mutations into the genome of influenza virus. Proc Natl Acad Sci U S A. 1990 May;87(10):3802–3805. doi: 10.1073/pnas.87.10.3802. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Enami M., Palese P. High-efficiency formation of influenza virus transfectants. J Virol. 1991 May;65(5):2711–2713. doi: 10.1128/jvi.65.5.2711-2713.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hausmann J., Kretzschmar E., Garten W., Klenk H. D. N1 neuraminidase of influenza virus A/FPV/Rostock/34 has haemadsorbing activity. J Gen Virol. 1995 Jul;76(Pt 7):1719–1728. doi: 10.1099/0022-1317-76-7-1719. [DOI] [PubMed] [Google Scholar]
- Horimoto T., Kawaoka Y. Reverse genetics provides direct evidence for a correlation of hemagglutinin cleavability and virulence of an avian influenza A virus. J Virol. 1994 May;68(5):3120–3128. doi: 10.1128/jvi.68.5.3120-3128.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huddleston J. A., Brownlee G. G. The sequence of the nucleoprotein gene of human influenza A virus, strain A/NT/60/68. Nucleic Acids Res. 1982 Feb 11;10(3):1029–1038. doi: 10.1093/nar/10.3.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ito T., Suzuki Y., Mitnaul L., Vines A., Kida H., Kawaoka Y. Receptor specificity of influenza A viruses correlates with the agglutination of erythrocytes from different animal species. Virology. 1997 Jan 20;227(2):493–499. doi: 10.1006/viro.1996.8323. [DOI] [PubMed] [Google Scholar]
- Laver W. G., Colman P. M., Webster R. G., Hinshaw V. S., Air G. M. Influenza virus neuraminidase with hemagglutinin activity. Virology. 1984 Sep;137(2):314–323. doi: 10.1016/0042-6822(84)90223-x. [DOI] [PubMed] [Google Scholar]
- Niwa H., Yamamura K., Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991 Dec 15;108(2):193–199. doi: 10.1016/0378-1119(91)90434-d. [DOI] [PubMed] [Google Scholar]
- Nuss J. M., Air G. M. Transfer of the hemagglutinin activity of influenza virus neuraminidase subtype N9 into an N2 neuraminidase background. Virology. 1991 Aug;183(2):496–504. doi: 10.1016/0042-6822(91)90979-l. [DOI] [PubMed] [Google Scholar]
- Palese P., Tobita K., Ueda M., Compans R. W. Characterization of temperature sensitive influenza virus mutants defective in neuraminidase. Virology. 1974 Oct;61(2):397–410. doi: 10.1016/0042-6822(74)90276-1. [DOI] [PubMed] [Google Scholar]
- Parvin J. D., Palese P., Honda A., Ishihama A., Krystal M. Promoter analysis of influenza virus RNA polymerase. J Virol. 1989 Dec;63(12):5142–5152. doi: 10.1128/jvi.63.12.5142-5152.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Suzuki Y., Matsunaga M., Matsumoto M. N-Acetylneuraminyllactosylceramide, GM3-NeuAc, a new influenza A virus receptor which mediates the adsorption-fusion process of viral infection. Binding specificity of influenza virus A/Aichi/2/68 (H3N2) to membrane-associated GM3 with different molecular species of sialic acid. J Biol Chem. 1985 Feb 10;260(3):1362–1365. [PubMed] [Google Scholar]
- Taylor J., Weinberg R., Kawaoka Y., Webster R. G., Paoletti E. Protective immunity against avian influenza induced by a fowlpox virus recombinant. Vaccine. 1988 Dec;6(6):504–508. doi: 10.1016/0264-410x(88)90101-6. [DOI] [PubMed] [Google Scholar]
- Varghese J. N., McKimm-Breschkin J. L., Caldwell J. B., Kortt A. A., Colman P. M. The structure of the complex between influenza virus neuraminidase and sialic acid, the viral receptor. Proteins. 1992 Nov;14(3):327–332. doi: 10.1002/prot.340140302. [DOI] [PubMed] [Google Scholar]
- WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
- Webster R. G., Air G. M., Metzger D. W., Colman P. M., Varghese J. N., Baker A. T., Laver W. G. Antigenic structure and variation in an influenza virus N9 neuraminidase. J Virol. 1987 Sep;61(9):2910–2916. doi: 10.1128/jvi.61.9.2910-2916.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Webster R. G., Hinshaw V. S., Laver W. G. Selection and analysis of antigenic variants of the neuraminidase of N2 influenza viruses with monoclonal antibodies. Virology. 1982 Feb;117(1):93–104. doi: 10.1016/0042-6822(82)90510-4. [DOI] [PubMed] [Google Scholar]