title: Membrane interactions of the tick-borne encephalitis virus fusion protein E at low pH.
abstract: Membrane fusion of the flavivirus tick-borne encephalitis virus is triggered by the mildly acidic pH of the endosome and is mediated by envelope protein E, a class II viral fusion protein. The low-pH trigger induces an oligomeric rearrangement in which the subunits of the native E homodimers dissociate and the monomeric subunits then reassociate into homotrimers. Here we provide evidence that membrane binding is mediated by the intermediate monomeric form of E, generated by low-pH-induced dissociation of the dimer. Liposome coflotation experiments revealed that association with target membranes occurred only when liposomes were present at the time of acidification, whereas pretreating virions at low pH in the absence of membranes resulted in the loss of their ability to stably attach to liposomes. With the cleavable cross-linker ethylene glycolbis(succinimidylsuccinate), it was shown that a truncated soluble form of the E protein (sE) could bind to membranes only when the dimers were free to dissociate at low pH, and binding could be blocked by a monoclonal antibody that recognizes the fusion peptide, which is at the distal tip of the E monomer but is buried in the native dimer. Surprisingly, analysis of the membrane-associated sE proteins revealed that they had formed trimers. This was unexpected because this protein lacks a sequence element in the C-terminal stem-anchor region, which was shown to be essential for trimerization in the absence of a target membrane. It can therefore be concluded that the formation of a trimeric form of sE is facilitated by membrane binding. Its stability is apparently maintained by contacts between the ectodomains only and is not dependent on sequence elements in the stem-anchor region as previously assumed.
authors.affiliation: Institute of Virology, University of Vienna, A-1095 Vienna, Austria. karin.stiasny@univie.ac.at
authors.forename: Karin
authors.initials: K
authors.lastname: Stiasny
authors.forename: Steven L
authors.initials: SL
authors.lastname: Allison
authors.forename: Juliane
authors.initials: J
authors.lastname: Schalich
authors.forename: Franz X
authors.initials: FX
authors.lastname: Heinz
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002642:Chick Embryo; D019281:Dimerization; D004669:Encephalitis Viruses, Tick-Borne / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism; D008561:Membrane Fusion; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism*; D014771:Virion / Q000378:metabolism
chemical_list: D008081:Liposomes; D014759:Viral Envelope Proteins; C065696:glycoprotein E, Flavivirus