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 Mahon, P. J.
Right arrow Articles by Iorio, R. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mahon, P. J.
Right arrow Articles by Iorio, R. M.

 Previous Article  |  Next Article 

Journal of Virology, November 2008, p. 10386-10396, Vol. 82, No. 21
0022-538X/08/$08.00+0     doi:10.1128/JVI.00581-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Engineered Intermonomeric Disulfide Bonds in the Globular Domain of Newcastle Disease Virus Hemagglutinin-Neuraminidase Protein: Implications for the Mechanism of Fusion Promotion{triangledown}

Paul J. Mahon,1,2 Anne M. Mirza,1 Thomas A. Musich,3 and Ronald M. Iorio1,3*

Department of Molecular Genetics and Microbiology,1 Program in Immunology and Virology, University of Massachusetts Medical School, Worcester, Massachusetts 01655,3 Department of Biology, Assumption College, Worcester, Massachusetts 016092

Received 14 March 2008/ Accepted 7 August 2008

The promotion of membrane fusion by Newcastle disease virus (NDV) requires an interaction between the viral hemagglutinin-neuraminidase (HN) and fusion (F) proteins, although the mechanism by which this interaction regulates fusion is not clear. The NDV HN protein exists as a tetramer composed of a pair of dimers. Based on X-ray crystallographic studies of the NDV HN globular domain (S. Crennell et al., Nat. Struct. Biol. 7:1068-1074, 2000), it was proposed that the protein undergoes a significant conformational change from an initial structure having minimal intermonomeric contacts to a structure with a much more extensive dimer interface. This conformational change was predicted to be integral to fusion promotion with the minimal interface form required to maintain F in its prefusion state until HN binds receptors. However, no evidence for such a conformational change exists for any other paramyxovirus attachment protein. To test the NDV model, we have engineered a pair of intermonomeric disulfide bonds across the dimer interface in the globular domain of an otherwise non-disulfide-linked NDV HN protein by the introduction of cysteine substitutions for residues T216 and D230. The disulfide-linked dimer is formed both intracellularly and in the absence of receptor binding and is efficiently expressed at the cell surface. The disulfide bonds preclude formation of the minimal interface form of the protein and yet enhance both receptor-binding activity at 37°C and fusion promotion. These results confirm that neither the minimal interface form of HN nor the proposed drastic conformational change in the protein is required for fusion.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655. Phone: (508) 856-5257. Fax: (508) 856-5920. E-mail: ronald.iorio{at}umassmed.edu

{triangledown} Published ahead of print on 27 August 2008.

Journal of Virology, November 2008, p. 10386-10396, Vol. 82, No. 21
0022-538X/08/$08.00+0     doi:10.1128/JVI.00581-08
Copyright © 2008, 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]  
  • Paal, T., Brindley, M. A., St. Clair, C., Prussia, A., Gaus, D., Krumm, S. A., Snyder, J. P., Plemper, R. K. (2009). Probing the Spatial Organization of Measles Virus Fusion Complexes. J. Virol. 83: 10480-10493 [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»