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 Porotto, M.
Right arrow Articles by Moscona, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Porotto, M.
Right arrow Articles by Moscona, A.

 Previous Article  |  Next Article 

Journal of Virology, March 2003, p. 3647-3654, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3647-3654.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Triggering of Human Parainfluenza Virus 3 Fusion Protein (F) by the Hemagglutinin-Neuraminidase (HN) Protein: an HN Mutation Diminishes the Rate of F Activation and Fusion

Matteo Porotto, Matthew Murrell, Olga Greengard, and Anne Moscona*

Department of Pediatrics, Mount Sinai School of Medicine, New York, New York 10029

Received 24 October 2002/ Accepted 13 December 2002

For human parainfluenza virus type 3 and many other paramyxoviruses, membrane fusion mediated by the fusion protein (F) has a stringent requirement for the presence of the homotypic hemagglutinin-neuraminidase protein (HN). With the goal of gaining further insight into the role of HN in the fusion process, we developed a simple method for quantitative comparison of the ability of wild-type and variant HNs to activate F. In this method, HN/F-coexpressing cells with red blood cells (RBC) bound to them at 4°C are transferred to 22°C, and at different times after transfer 4-guanidino-neu5Ac2en (4-GU-DANA) is added; this inhibitor of the HN-receptor interaction then releases all reversibly bound RBC but not those in which F insertion in the target membrane or fusion has occurred. Thus, the amount of irreversibly bound (nonreleased) RBC provides a measure of F activation, and the use of fluorescently labeled RBC permits microscopic assessment of the extent to which F insertion has progressed to fusion. We studied two neuraminidase-deficient HN variants, C28a, which has two mutations, P111S and D216N, and C28, which possesses the D216N mutation only. C28a but not C28 exhibits a slow fusion phenotype, although determination of the HNs' receptor-binding avidity (with our sensitive method, employing RBC with different degrees of receptor depletion) showed that the receptor-binding avidity of C28a or C28 HN was not lower than that of the wild type. The F activation assay, however, revealed fusion-triggering defects in C28a HN. After 10 and also 20 min at 22°C, irreversible RBC binding was significantly less for cells coexpressing wild-type F with C28a HN than for cells coexpressing wild-type F with wild-type HN. In addition, F insertion progressed to fusion more slowly in the case of C28a HN-expressing cells than of wild-type HN-expressing cells. Identical defects were found for P111S HN, whereas for C28 HN, representing the 216 mutation of C28a, F activation and fusion were as rapid as for wild-type HN. The diminished fusion promotion capacity of C28a HN is therefore attributable to P111S, a mutation in the stalk region of the molecule that causes no decrease in receptor-binding avidity. C28a HN is the first parainfluenza virus variant found so far to be specifically defective in HN's F-triggering and fusion promotion functions and may contribute to our understanding of transmission of the activating signal from HN to F.

* Corresponding author. Mailing address: Department of Pediatrics, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029. Phone: (212) 241-9709. Fax: (212) 860-3316. E-mail: Anne.moscona{at}mssm.edu.

Journal of Virology, March 2003, p. 3647-3654, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3647-3654.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Connolly, S. A., Leser, G. P., Jardetzky, T. S., Lamb, R. A. (2009). Bimolecular Complementation of Paramyxovirus Fusion and Hemagglutinin-Neuraminidase Proteins Enhances Fusion: Implications for the Mechanism of Fusion Triggering. J. Virol. 83: 10857-10868 [Abstract] [Full Text]  
  • Porotto, M., Yokoyama, C. C., Orefice, G., Kim, H.-S., Aljofan, M., Mungall, B. A., Moscona, A. (2009). Kinetic Dependence of Paramyxovirus Entry Inhibition. J. Virol. 83: 6947-6951 [Abstract] [Full Text]  
  • Porotto, M., Orefice, G., Yokoyama, C. C., Mungall, B. A., Realubit, R., Sganga, M. L., Aljofan, M., Whitt, M., Glickman, F., Moscona, A. (2009). Simulating Henipavirus Multicycle Replication in a Screening Assay Leads to Identification of a Promising Candidate for Therapy. J. Virol. 83: 5148-5155 [Abstract] [Full Text]  
  • Aguilar, H. C., Ataman, Z. A., Aspericueta, V., Fang, A. Q., Stroud, M., Negrete, O. A., Kammerer, R. A., Lee, B. (2009). A Novel Receptor-induced Activation Site in the Nipah Virus Attachment Glycoprotein (G) Involved in Triggering the Fusion Glycoprotein (F). J. Biol. Chem. 284: 1628-1635 [Abstract] [Full Text]  
  • Jain, S., McGinnes, L. W., Morrison, T. G. (2009). Role of Thiol/Disulfide Exchange in Newcastle Disease Virus Entry. J. Virol. 83: 241-249 [Abstract] [Full Text]  
  • Tsurudome, M., Nishio, M., Ito, M., Tanahashi, S., Kawano, M., Komada, H., Ito, Y. (2008). Effects of Hemagglutinin-Neuraminidase Protein Mutations on Cell-Cell Fusion Mediated by Human Parainfluenza Type 2 Virus. J. Virol. 82: 8283-8295 [Abstract] [Full Text]  
  • Porotto, M., Carta, P., Deng, Y., Kellogg, G. E., Whitt, M., Lu, M., Mungall, B. A., Moscona, A. (2007). Molecular Determinants of Antiviral Potency of Paramyxovirus Entry Inhibitors. J. Virol. 81: 10567-10574 [Abstract] [Full Text]  
  • Corey, E. A., Iorio, R. M. (2007). Mutations in the Stalk of the Measles Virus Hemagglutinin Protein Decrease Fusion but Do Not Interfere with Virus-Specific Interaction with the Homologous Fusion Protein. J. Virol. 81: 9900-9910 [Abstract] [Full Text]  
  • Palermo, L. M., Porotto, M., Greengard, O., Moscona, A. (2007). Fusion Promotion by a Paramyxovirus Hemagglutinin-Neuraminidase Protein: pH Modulation of Receptor Avidity of Binding Sites I and II. J. Virol. 81: 9152-9161 [Abstract] [Full Text]  
  • Porotto, M., Fornabaio, M., Kellogg, G. E., Moscona, A. (2007). A Second Receptor Binding Site on Human Parainfluenza Virus Type 3 Hemagglutinin-Neuraminidase Contributes to Activation of the Fusion Mechanism. J. Virol. 81: 3216-3228 [Abstract] [Full Text]  
  • Cantin, C., Holguera, J., Ferreira, L., Villar, E., Munoz-Barroso, I. (2007). Newcastle disease virus may enter cells by caveolae-mediated endocytosis. J. Gen. Virol. 88: 559-569 [Abstract] [Full Text]  
  • Porotto, M., Doctor, L., Carta, P., Fornabaio, M., Greengard, O., Kellogg, G. E., Moscona, A. (2006). Inhibition of Hendra Virus Fusion. J. Virol. 80: 9837-9849 [Abstract] [Full Text]  
  • Bousse, T., Takimoto, T. (2006). Mutation at residue 523 creates a second receptor binding site on human parainfluenza virus type 1 hemagglutinin-neuraminidase protein.. J. Virol. 80: 9009-9016 [Abstract] [Full Text]  
  • McGinnes, L. W., Morrison, T. G. (2006). Inhibition of Receptor Binding Stabilizes Newcastle Disease Virus HN and F Protein-Containing Complexes. J. Virol. 80: 2894-2903 [Abstract] [Full Text]  
  • Porotto, M., Fornabaio, M., Greengard, O., Murrell, M. T., Kellogg, G. E., Moscona, A. (2006). Paramyxovirus Receptor-Binding Molecules: Engagement of One Site on the Hemagglutinin-Neuraminidase Protein Modulates Activity at the Second Site. J. Virol. 80: 1204-1213 [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]  
  • Russell, C. J., Jardetzky, T. S., Lamb, R. A. (2004). Conserved Glycine Residues in the Fusion Peptide of the Paramyxovirus Fusion Protein Regulate Activation of the Native State. J. Virol. 78: 13727-13742 [Abstract] [Full Text]  
  • Porotto, M., Murrell, M., Greengard, O., Lawrence, M. C., McKimm-Breschkin, J. L., Moscona, A. (2004). Inhibition of Parainfluenza Virus Type 3 and Newcastle Disease Virus Hemagglutinin-Neuraminidase Receptor Binding: Effect of Receptor Avidity and Steric Hindrance at the Inhibitor Binding Sites. J. Virol. 78: 13911-13919 [Abstract] [Full Text]  
  • Melanson, V. R., Iorio, R. M. (2004). Amino Acid Substitutions in the F-Specific Domain in the Stalk of the Newcastle Disease Virus HN Protein Modulate Fusion and Interfere with Its Interaction with the F Protein. J. Virol. 78: 13053-13061 [Abstract] [Full Text]  
  • Meyerholz, D. K., Grubor, B., Gallup, J. M., Lehmkuhl, H. D., Anderson, R. D., Lazic, T., Ackermann, M. R. (2004). Adenovirus-Mediated Gene Therapy Enhances Parainfluenza Virus 3 Infection in Neonatal Lambs. J. Clin. Microbiol. 42: 4780-4787 [Abstract] [Full Text]  
  • Li, J., Quinlan, E., Mirza, A., Iorio, R. M. (2004). Mutated Form of the Newcastle Disease Virus Hemagglutinin-Neuraminidase Interacts with the Homologous Fusion Protein despite Deficiencies in both Receptor Recognition and Fusion Promotion. J. Virol. 78: 5299-5310 [Abstract] [Full Text]  
  • Gravel, K. A., Morrison, T. G. (2003). Interacting Domains of the HN and F Proteins of Newcastle Disease Virus. J. Virol. 77: 11040-11049 [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»