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

 Previous Article  |  Next Article 

J. Virol., 05 1997, 3719-3725, Vol 71, No. 5
Copyright © 1997, American Society for Microbiology

Oligosaccharides in the stem region maintain the influenza virus hemagglutinin in the metastable form required for fusion activity

R Ohuchi, M Ohuchi, W Garten and HD Klenk
Institute fur Virologie, Philipps-Universitat Marburg, Germany.

The influenza A virus hemagglutinin (HA) has three conserved oligosaccharides located in the stem region at asparagine residues 12, 28, and 478. The biological role of these oligosaccharides has been investigated by mutational analysis of HA of fowl plague virus that was expressed from a simian virus 40 vector in the presence of ammonium chloride for protection from acid denaturation in the trans-Golgi network. Resistance to endoglycosidase H and cleavage of HA into the subunits HA1 and HA2 have been analyzed as markers for intracellular transport. Cell surface exposure has been determined by hemadsorption following neuraminidase treatment, by immunofluorescence staining, and by fluorescence-activated cell sorter analysis. When all three stem oligosaccharides were removed, transport was almost completely blocked. When two of the three stem oligosaccharides, particularly those at asparagine residues 12 and 28, were missing, HA was transported to the surface but showed extremely low fusion activity. With mutants lacking one stem oligosaccharide, fusion was reduced to a lesser extent. Removal of stem oligosaccharides resulted also in an increase in the pH optimum required for fusion. On the other hand, no reduction in fusion activity was observed when oligosaccharides in the head region of the HA spike were removed. These results indicate that the conserved oligosaccharides in the stem stabilize HA in the form susceptible to the conformational change necessary for fusion.

This article has been cited by other articles:

• Helle, F., Goffard, A., Morel, V., Duverlie, G., McKeating, J., Keck, Z.-Y., Foung, S., Penin, F., Dubuisson, J., Voisset, C. (2007). The Neutralizing Activity of Anti-Hepatitis C Virus Antibodies Is Modulated by Specific Glycans on the E2 Envelope Protein. J. Virol. 81: 8101-8111 [Abstract] [Full Text]  
• Dowling, W., Thompson, E., Badger, C., Mellquist, J. L., Garrison, A. R., Smith, J. M., Paragas, J., Hogan, R. J., Schmaljohn, C. (2007). Influences of Glycosylation on Antigenicity, Immunogenicity, and Protective Efficacy of Ebola Virus GP DNA Vaccines. J. Virol. 81: 1821-1837 [Abstract] [Full Text]  
• Goffard, A., Callens, N., Bartosch, B., Wychowski, C., Cosset, F.-L., Montpellier, C., Dubuisson, J. (2005). Role of N-Linked Glycans in the Functions of Hepatitis C Virus Envelope Glycoproteins. J. Virol. 79: 8400-8409 [Abstract] [Full Text]  
• Abe, Y., Takashita, E., Sugawara, K., Matsuzaki, Y., Muraki, Y., Hongo, S. (2004). Effect of the Addition of Oligosaccharides on the Biological Activities and Antigenicity of Influenza A/H3N2 Virus Hemagglutinin. J. Virol. 78: 9605-9611 [Abstract] [Full Text]  
• von Messling, V., Milosevic, D., Devaux, P., Cattaneo, R. (2004). Canine Distemper Virus and Measles Virus Fusion Glycoprotein Trimers: Partial Membrane-Proximal Ectodomain Cleavage Enhances Function. J. Virol. 78: 7894-7903 [Abstract] [Full Text]  
• Panda, A., Elankumaran, S., Krishnamurthy, S., Huang, Z., Samal, S. K. (2004). Loss of N-Linked Glycosylation from the Hemagglutinin- Neuraminidase Protein Alters Virulence of Newcastle Disease Virus. J. Virol. 78: 4965-4975 [Abstract] [Full Text]  
• von Messling, V., Cattaneo, R. (2003). N-Linked Glycans with Similar Location in the Fusion Protein Head Modulate Paramyxovirus Fusion. J. Virol. 77: 10202-10212 [Abstract] [Full Text]  
• Tsuchiya, E., Sugawara, K., Hongo, S., Matsuzaki, Y., Muraki, Y., Nakamura, K. (2002). Role of overlapping glycosylation sequons in antigenic properties, intracellular transport and biological activities of influenza A/H2N2 virus haemagglutinin. J. Gen. Virol. 83: 3067-3074 [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]  
• 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]  
• 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]