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Journal of Virology, March 2000, p. 2714-2720, Vol. 74, No. 6
Department of Biophysical Chemistry,
Biozentrum of the University of Basel, CH 4056 Basel,
Switzerland,1 Institut de Pharmacologie
et de Biologie Structurale, CNRS UPR 9062, 31077 Toulouse Cedex,
France,3 and Laboratory of Molecular Virology,
Department of Medical Microbiology, University of Groningen,
9713 AV Groningen, The Netherlands2
Received 16 August 1999/Accepted 14 December 1999
Membrane fusion mediated by influenza virus hemagglutinin (HA) is
believed to proceed via the cooperative action of multiple HA trimers.
To determine the minimal number of HA trimers required to trigger
fusion, and to assess the importance of cooperativity between these HA
trimers, we have generated virosomes containing coreconstituted HAs
derived from two strains of virus with different pH dependencies for
fusion, X-47 (optimal fusion at pH 5.1; threshold at pH 5.6) and
A/Shangdong (optimal fusion at pH 5.6; threshold at pH 6.0), and
measured fusion of these virosomes with erythrocyte ghosts by a
fluorescence lipid mixing assay. Virosomes with different X-47-to-A/Shangdong HA ratios, at a constant HA-to-lipid ratio, showed
comparable ghost-binding activities, and the low-pH-induced conformational change of A/Shangdong HA did not affect the fusion activity of X-47 HA. The initial rate of fusion of these virosomes at
pH 5.7 increased directly proportional to the surface density of
A/Shangdong HA, and a single A/Shangdong trimer per virosome appeared
to suffice to induce fusion. The reciprocal of the lag time before the
onset of fusion was directly proportional to the surface density of
fusion-competent HA. These results support the notion that there is no
cooperativity between HA trimers during influenza virus fusion.
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Role of Hemagglutinin Surface Density in the
Initial Stages of Influenza Virus Fusion: Lack of Evidence for
Cooperativity


*
Corresponding author. Mailing address for Jan Wilschut:
Department of Medical Microbiology, University of Groningen, Ant. Deusinglaan 1, 9713 AV Groningen, The Netherlands. Phone: 31 50 3632733. Fax: 31 50 3632728. E-mail: J.C.Wilschut{at}med.rug.nl. Mailing
address for Toon Stegmann: Institut de Pharmacologie et de Biologie
Structurale, CNRS UPR 9062, 205 Route de Narbonne, 31077 Toulouse
Cedex, France. Phone: 33 5 61 17 54 63. Fax: 33 5 61 17 59 94. E-mail:
stegmann{at}ipbs.fr.
Present address: s_ausborn{at}hotmail.com.
Present address: Haemoprobe BV, 9713 GX Groningen, The Netherlands.
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