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Single-particle kinetics of influenza virus membrane fusion

Journal Article


Abstract


  • Membrane fusion is an essential step during entry of enveloped viruses into cells. Conventional fusion assays are generally limited to observation of ensembles of multiple fusion events, confounding more detailed analysis of the sequence of the molecular steps involved. We have developed an in vitro, two-color fluorescence assay to monitor kinetics of single virus particles fusing with a target bilayer on an essentially fluid support. Analysis of lipid- and content-mixing trajectories on a particle-by-particle basis provides evidence for multiple, long-lived kinetic intermediates leading to hemifusion, followed by a single, rate-limiting step to pore formation. We interpret the series of intermediates preceding hemifusion as a result of the requirement that multiple copies of the trimeric hemagglutinin fusion protein be activated to initiate the fusion process.

Authors


  •   Floyd, Daniel L. (external author)
  •   Ragains, Justin R. (external author)
  •   Skehel, John J. (external author)
  •   Harrison, Stephen C. (external author)
  •   van Oijen, Antoine M.

Publication Date


  • 2008

Citation


  • Floyd, D. L., Ragains, J. R., Skehel, J. J., Harrison, S. C. & van Oijen, A. M. (2008). Single-particle kinetics of influenza virus membrane fusion. Proceedings of the National Academy of Sciences of USA, 105 (40), 15382-15387.

Scopus Eid


  • 2-s2.0-55749106791

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2180

Has Global Citation Frequency


Number Of Pages


  • 5

Start Page


  • 15382

End Page


  • 15387

Volume


  • 105

Issue


  • 40

Place Of Publication


  • United States

Abstract


  • Membrane fusion is an essential step during entry of enveloped viruses into cells. Conventional fusion assays are generally limited to observation of ensembles of multiple fusion events, confounding more detailed analysis of the sequence of the molecular steps involved. We have developed an in vitro, two-color fluorescence assay to monitor kinetics of single virus particles fusing with a target bilayer on an essentially fluid support. Analysis of lipid- and content-mixing trajectories on a particle-by-particle basis provides evidence for multiple, long-lived kinetic intermediates leading to hemifusion, followed by a single, rate-limiting step to pore formation. We interpret the series of intermediates preceding hemifusion as a result of the requirement that multiple copies of the trimeric hemagglutinin fusion protein be activated to initiate the fusion process.

Authors


  •   Floyd, Daniel L. (external author)
  •   Ragains, Justin R. (external author)
  •   Skehel, John J. (external author)
  •   Harrison, Stephen C. (external author)
  •   van Oijen, Antoine M.

Publication Date


  • 2008

Citation


  • Floyd, D. L., Ragains, J. R., Skehel, J. J., Harrison, S. C. & van Oijen, A. M. (2008). Single-particle kinetics of influenza virus membrane fusion. Proceedings of the National Academy of Sciences of USA, 105 (40), 15382-15387.

Scopus Eid


  • 2-s2.0-55749106791

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2180

Has Global Citation Frequency


Number Of Pages


  • 5

Start Page


  • 15382

End Page


  • 15387

Volume


  • 105

Issue


  • 40

Place Of Publication


  • United States