Dilation of the influenza hemagglutinin fusion pore revealed by the kinetics of individual cell-cell fusion events

doi:10.1083/jcb.135.1.63

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Dilation of the influenza hemagglutinin fusion pore revealed by the kinetics of individual cell-cell fusion events

R Blumenthal, DP Sarkar, S Durell, DE Howard and SJ Morris
Section on Membrane Structure and Function, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. blumen@helix.nih.gov

We have monitored kinetics of fusion between cell pairs consisting of a single influenza hemaglutinin (HA)-expressing cell and a single erythrocyte (RBC) that had been labeled with both a fluorescent lipid (Dil) in the membrane and a fluorescent solute (calcein) in the aqueous space. Initial fusion pore opening between the RBC and HA-expressing cell produced a change in RBC membrane potential (delta psi) that was monitored by a decrease in Dil fluorescence. This event was followed by two distinct stages of fusion pore dilation: the flux of fluorescent lipid (phi L) and the flux of a large aqueous fluorescent dye (phi s). We have analyzed the kinetics of events that occur as a result of transitions between a fusion pore (FP) and a solute permissive fusion pore (FPs). Our data are consistent with a fusion pore comprising six HA trimers.
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