Intermediates in influenza induced membrane fusion.
Our results show that the mechanism by which influenza virus fuses with target membranes involves sequential complex changes in the hemagglutinin (HA, the viral fusion protein) and in the contact site between virus and target membrane. To render individual steps amenable to study, we worked at 0 degree C which decreased the rate of fusion and increased the efficiency. The mechanism of fusion at 0 degree C and 37 degrees C was similar. The process began with a conformational change in HA which exposed the fusion peptides but did not lead to dissociation of the tops of the ectodomain of the trimer. The change in the protein led to immediate hydrophobic attachment of the virus to the target liposomes. Attachment was followed by a lag period (4-8 min at 0 degree C, 0.6-2 s at 37 degrees C) during which rearrangements occurred in the site of membrane contact between the virus and liposome. After a further series of changes the final bilayer merger took place. This final fusion event was not pH dependent. At 0 degree C efficient fusion occurred without dissociation of the top domains of the HA trimer, suggesting that a transient conformation of HA is responsible for fusion at physiological temperatures. The observations lead to a revised model for HA mediated fusion.
Stegmann
T
T
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510.
White
J M
JM
Helenius
A
A
D000818:Animals; D002417:Cattle; D003588:Cytopathogenic Effect, Viral; D004910:Erythrocyte Membrane / Q000502:physiology; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008561:Membrane Fusion; D009975:Orthomyxoviridae / Q000254:growth & development / Q000502:physiology*; D011487:Protein Conformation; D011991:Receptors, Virus / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013696:Temperature; D014759:Viral Envelope Proteins / Q000737:chemistry*; D014775:Virus Activation
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D008081:Liposomes; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins