This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wagner, R.
Right arrow Articles by Klenk, H.-D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wagner, R.
Right arrow Articles by Klenk, H.-D.

 Previous Article  |  Next Article 

Journal of Virology, July 2000, p. 6316-6323, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Interdependence of Hemagglutinin Glycosylation and Neuraminidase as Regulators of Influenza Virus Growth: a Study by Reverse Genetics

Ralf Wagner,1 Thorsten Wolff,1,dagger Astrid Herwig,1 Stephan Pleschka,2 and Hans-Dieter Klenk1,*

Institut für Virologie, Philipps-Universität, 35011 Marburg,1 and Institut für Mikrobiologie und Molekularbiologie, Justus Liebig-Universität Giessen, 35392 Giessen,2 Germany

Received 15 December 1999/Accepted 25 April 2000

The hemagglutinin (HA) of fowl plague virus A/FPV/Rostock/34 (H7N1) carries two N-linked oligosaccharides attached to Asn123 and Asn149 in close vicinity to the receptor-binding pocket. In previous studies in which HA mutants lacking either one (mutants G1 and G2) or both (mutant G1,2) glycosylation sites had been expressed from a simian virus 40 vector, we showed that these glycans regulate receptor binding affinity (M. Ohuchi, R. Ohuchi, A. Feldmann, and H. D. Klenk, J. Virol. 71:8377-8384, 1997). We have now investigated the effect of these mutations on virus growth using recombinant viruses generated by an RNA polymerase I-based reverse genetics system. Two reassortants of influenza virus strain A/WSN/33 were used as helper viruses to obtain two series of HA mutant viruses differing only in the neuraminidase (NA). Studies using N1 NA viruses revealed that loss of the oligosaccharide from Asn149 (mutant G2) or loss of both oligosaccharides (mutant G1,2) has a pronounced effect on virus growth in MDCK cells. Growth of virus lacking both oligosaccharides from infected cells was retarded, and virus yields in the medium were decreased about 20-fold. Likewise, there was a reduction in plaque size that was distinct with G1,2 and less pronounced with G2. These effects could be attributed to a highly impaired release of mutant progeny viruses from host cells. In contrast, with recombinant viruses containing N2 NA, these restrictions were much less apparent. N1 recombinants showed lower neuraminidase activity than N2 recombinants, indicating that N2 NA is able to partly overrule the high-affinity binding of mutant HA to the receptor. These results demonstrate that N-glycans flanking the receptor-binding site of the HA molecule are potent regulators of influenza virus growth, with the glycan at Asn149 being dominant and that at Asn123 being less effective. In addition, we show here that HA and NA activities need to be highly balanced in order to allow productive influenza virus infection.

* Corresponding author. Mailing address: Institut für Virologie, Philipps-Universität Marburg, Postfach 2360, 35011 Marburg, Germany. Phone: 49/6421/28-66253. Fax: 49/6421/28-68962. E-mail: Klenk{at}mailer.uni-marburg.de.

dagger Present address: Robert-Koch-Institut, 13353 Berlin, Germany.

Journal of Virology, July 2000, p. 6316-6323, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Matsuoka, Y., Swayne, D. E., Thomas, C., Rameix-Welti, M.-A., Naffakh, N., Warnes, C., Altholtz, M., Donis, R., Subbarao, K. (2009). Neuraminidase Stalk Length and Additional Glycosylation of the Hemagglutinin Influence the Virulence of Influenza H5N1 Viruses for Mice. J. Virol. 83: 4704-4708 [Abstract] [Full Text]  
  • McDonald, N. J., Smith, C. B., Cox, N. J. (2007). Antigenic drift in the evolution of H1N1 influenza A viruses resulting from deletion of a single amino acid in the haemagglutinin gene. J. Gen. Virol. 88: 3209-3213 [Abstract] [Full Text]  
  • Vigerust, D. J., Ulett, K. B., Boyd, K. L., Madsen, J., Hawgood, S., McCullers, J. A. (2007). N-Linked Glycosylation Attenuates H3N2 Influenza Viruses. J. Virol. 81: 8593-8600 [Abstract] [Full Text]  
  • Ehrhardt, C., Wolff, T., Pleschka, S., Planz, O., Beermann, W., Bode, J. G., Schmolke, M., Ludwig, S. (2007). Influenza A Virus NS1 Protein Activates the PI3K/Akt Pathway To Mediate Antiapoptotic Signaling Responses. J. Virol. 81: 3058-3067 [Abstract] [Full Text]  
  • Long, G., Pan, X., Vlak, J. M. (2007). Absence of N-linked glycans from the F2 subunit of the major baculovirus envelope fusion protein F enhances fusogenicity. J. Gen. Virol. 88: 441-449 [Abstract] [Full Text]  
  • Landolt, G. A., Karasin, A. I., Schutten, M. M., Olsen, C. W. (2006). Restricted Infectivity of a Human-Lineage H3N2 Influenza A Virus in Pigs Is Hemagglutinin and Neuraminidase Gene Dependent. J. Clin. Microbiol. 44: 297-301 [Abstract] [Full Text]  
  • Giannecchini, S., Campitelli, L., Calzoletti, L., De Marco, M. A., Azzi, A., Donatelli, I. (2006). Comparison of in vitro replication features of H7N3 influenza viruses from wild ducks and turkeys: potential implications for interspecies transmission. J. Gen. Virol. 87: 171-175 [Abstract] [Full Text]  
  • Mishin, V. P., Novikov, D., Hayden, F. G., Gubareva, L. V. (2005). Effect of Hemagglutinin Glycosylation on Influenza Virus Susceptibility to Neuraminidase Inhibitors. J. Virol. 79: 12416-12424 [Abstract] [Full Text]  
  • Suzuki, T., Takahashi, T., Guo, C.-T., Hidari, K. I.-P. J., Miyamoto, D., Goto, H., Kawaoka, Y., Suzuki, Y. (2005). Sialidase Activity of Influenza A Virus in an Endocytic Pathway Enhances Viral Replication. J. Virol. 79: 11705-11715 [Abstract] [Full Text]  
  • Lu, B., Zhou, H., Ye, D., Kemble, G., Jin, H. (2005). Improvement of Influenza A/Fujian/411/02 (H3N2) Virus Growth in Embryonated Chicken Eggs by Balancing the Hemagglutinin and Neuraminidase Activities, Using Reverse Genetics. J. Virol. 79: 6763-6771 [Abstract] [Full Text]  
  • Porotto, M., Murrell, M., Greengard, O., Doctor, L., Moscona, A. (2005). Influence of the Human Parainfluenza Virus 3 Attachment Protein's Neuraminidase Activity on Its Capacity To Activate the Fusion Protein. J. Virol. 79: 2383-2392 [Abstract] [Full Text]  
  • Hulse, D. J., Webster, R. G., Russell, R. J., Perez, D. R. (2004). Molecular Determinants within the Surface Proteins Involved in the Pathogenicity of H5N1 Influenza Viruses in Chickens. J. Virol. 78: 9954-9964 [Abstract] [Full Text]  
  • Nimmerjahn, F., Dudziak, D., Dirmeier, U., Hobom, G., Riedel, A., Schlee, M., Staudt, L. M., Rosenwald, A., Behrends, U., Bornkamm, G. W., Mautner, J. (2004). Active NF-{kappa}B signalling is a prerequisite for influenza virus infection. J. Gen. Virol. 85: 2347-2356 [Abstract] [Full Text]  
  • Moll, M., Kaufmann, A., Maisner, A. (2004). Influence of N-Glycans on Processing and Biological Activity of the Nipah Virus Fusion Protein. J. Virol. 78: 7274-7278 [Abstract] [Full Text]  
  • Thompson, C. I., Barclay, W. S., Zambon, M. C. (2004). Changes in in vitro susceptibility of influenza A H3N2 viruses to a neuraminidase inhibitor drug during evolution in the human host. J Antimicrob Chemother 53: 759-765 [Abstract] [Full Text]  
  • Spackman, E., Senne, D. A., Davison, S., Suarez, D. L. (2003). Sequence Analysis of Recent H7 Avian Influenza Viruses Associated with Three Different Outbreaks in Commercial Poultry in the United States. J. Virol. 77: 13399-13402 [Abstract] [Full Text]  
  • Watanabe, T., Watanabe, S., Noda, T., Fujii, Y., Kawaoka, Y. (2003). Exploitation of Nucleic Acid Packaging Signals To Generate a Novel Influenza Virus-Based Vector Stably Expressing Two Foreign Genes. J. Virol. 77: 10575-10583 [Abstract] [Full Text]  
  • Flandorfer, A., Garcia-Sastre, A., Basler, C. F., Palese, P. (2003). Chimeric Influenza A Viruses with a Functional Influenza B Virus Neuraminidase or Hemagglutinin. J. Virol. 77: 9116-9123 [Abstract] [Full Text]  
  • Ohuchi, M., Ohuchi, R., Sakai, T., Matsumoto, A. (2002). Tight Binding of Influenza Virus Hemagglutinin to Its Receptor Interferes with Fusion Pore Dilation. J. Virol. 76: 12405-12413 [Abstract] [Full Text]  
  • Gubareva, L. V., Nedyalkova, M. S., Novikov, D. V., Murti, K. G., Hoffmann, E., Hayden, F. G. (2002). A release-competent influenza A virus mutant lacking the coding capacity for the neuraminidase active site. J. Gen. Virol. 83: 2683-2692 [Abstract] [Full Text]  
  • Sandrin, V., Boson, B., Salmon, P., Gay, W., Negre, D., Le Grand, R., Trono, D., Cosset, F.-L. (2002). Lentiviral vectors pseudotyped with a modified RD114 envelope glycoprotein show increased stability in sera and augmented transduction of primary lymphocytes and CD34+ cells derived from human and nonhuman primates. Blood 100: 823-832 [Abstract] [Full Text]  
  • Tsuchiya, E., Sugawara, K., Hongo, S., Matsuzaki, Y., Muraki, Y., Li, Z.-N., Nakamura, K. (2002). Effect of addition of new oligosaccharide chains to the globular head of influenza A/H2N2 virus haemagglutinin on the intracellular transport and biological activities of the molecule. J. Gen. Virol. 83: 1137-1146 [Abstract] [Full Text]  
  • Wagner, R., Heuer, D., Wolff, T., Herwig, A., Klenk, H.-D. (2002). N-Glycans attached to the stem domain of haemagglutinin efficiently regulate influenza A virus replication. J. Gen. Virol. 83: 601-609 [Abstract] [Full Text]  
  • Porotto, M., Greengard, O., Poltoratskaia, N., Horga, M.-A., Moscona, A. (2001). Human Parainfluenza Virus Type 3 HN-Receptor Interaction: Effect of 4-Guanidino-Neu5Ac2en on a Neuraminidase-Deficient Variant. J. Virol. 75: 7481-7488 [Abstract] [Full Text]  
  • Zimmer, G., Trotz, I., Herrler, G. (2001). N-Glycans of F Protein Differentially Affect Fusion Activity of Human Respiratory Syncytial Virus. J. Virol. 75: 4744-4751 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»