title: The role of target membrane sialic acid residues in the fusion activity of the influenza virus: the effect of two types of ganglioside on the kinetics of membrane merging.
abstract: The influenza virus enters target cells via the action of hemagglutinin proteins (HA) inserted into the viral envelope. HA promotes membrane fusion between the viral envelope and endosomal membrane at low pH, following viral binding to sialic acid-containing receptors on target cells, and internalization by endocytosis. The effect of target membrane sialic acid residues on the fusion activity of the influenza virus towards model membranes was evaluated by both reduction, (i.e. treating somatic cells with neuraminidase- (NA-) prior to virus-cell interactions), and by supplementing liposomes with the gangliosides GD1a and GT1b. The harshness of the neuraminidase pretreatment of target cells required to affect virus-induced membrane merging was found to greatly depend on the assay conditions, i.e. whether a virus-cell prebinding step at neutral pH was included prior to acidification. Minor concentrations of neuraminidase were found to greatly reduce virus fusion, but only in the absence of a prebinding step; they had no effect if this step was included. Although membrane merging was greatly reduced following cell neuraminidase pretreatment, virus-cell association at low pH was not disturbed proportionately. This probably reflects unspecific virus-cell binding under these conditions, probably of inactivated or aggregated virus particles, which does not translate into membrane merging. This seems to suggest both that target membrane sialic acid can protect the virus from losing its activity before triggering membrane merging, and that the importance of this interaction is not merely to ensure virus-target proximity. With liposomes, we found that both types of ganglioside supported efficient fusion, with GD1a promoting a slightly faster initial rate. However, in this case, virus-target proximity closely mirrored fusion activity, thus pointing to differential specificity between targets routinely used to assay influenza virus fusion activity.
authors.affiliation: Center for Neuroscience and Cell Biology, University of Coimbra, Portugal. jramalho@ci.uc.pt
authors.forename: João
authors.initials: J
authors.lastname: Ramalho-Santos
authors.forename: Maria C
authors.initials: MC
authors.lastname: Pedroso De Lima
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D002642:Chick Embryo; D005732:Gangliosides / Q000737:chemistry / Q000378:metabolism*; D006389:Hemagglutinins, Viral / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008561:Membrane Fusion / Q000502:physiology*; D009439:Neuraminidase / Q000378:metabolism; D009975:Orthomyxoviridae / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism*
chemical_list: D005732:Gangliosides; D006389:Hemagglutinins, Viral; D008081:Liposomes; D021122:Protein Subunits; D012794:Sialic Acids; D009439:Neuraminidase