title: Recombinant subviral particles from tick-borne encephalitis virus are fusogenic and provide a model system for studying flavivirus envelope glycoprotein functions.
abstract: Recombinant subviral particles (RSPs) obtained by coexpression of the envelope (E) and premembrane (prM) proteins of tick-borne encephalitis virus in COS cells (S. L. Allison, K. Stadler, C. W. Mandl, C. Kunz, and F. X. Heinz, J. Virol. 69:5816-5820, 1995) were extensively characterized and shown to be ordered structures containing envelope glycoproteins with structural and functional properties very similar to those in the virion envelope. The particles were spherical, with a diameter of about 30 nm and a buoyant density of 1.14 g/cm3 in sucrose gradients. They contained mature E proteins with endoglycosidase H-resistant glycans as well as fully cleaved mature M proteins. Cleavage of prM, which requires an acidic pH in exocytic compartments, could be inhibited by treatment of transfected cells with ammonium chloride, implying a common maturation pathway for RSPs and virions. RSPs incorporated [14C]choline but not [3H]uridine, demonstrating that they contain lipid but probably lack nucleic acid. The envelope proteins of RSPs exhibited a native antigenic and oligomeric structure compared with virions, and incubation at an acidic pH (pH <6.5) induced identical conformational changes and structural rearrangements, including an irreversible quantitative conversion of dimers to trimers. The RSPs were also shown to be functionally active, inducing membrane fusion in a low-pH-dependent manner and demonstrating the same specific hemagglutination activity as whole virions. Tick-borne encephalitis virus RSPs thus represent an excellent model system for investigating the structural basis of viral envelope glycoprotein functions.
authors.affiliation: Institute of Virology, University of Vienna, Austria.
authors.forename: J
authors.initials: J
authors.lastname: Schalich
authors.forename: S L
authors.initials: SL
authors.lastname: Allison
authors.forename: K
authors.initials: K
authors.lastname: Stiasny
authors.forename: C W
authors.initials: CW
authors.lastname: Mandl
authors.forename: C
authors.initials: C
authors.lastname: Kunz
authors.forename: F X
authors.initials: FX
authors.lastname: Heinz
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D004279:DNA, Viral; D004669:Encephalitis Viruses, Tick-Borne / Q000737:chemistry / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D006384:Hemagglutination; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000032:analysis; D008954:Models, Biological; D009696:Nucleic Acids / Q000032:analysis; D011487:Protein Conformation; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000502:physiology*; D014771:Virion / Q000737:chemistry / Q000235:genetics / Q000502:physiology
chemical_list: D004279:DNA, Viral; D008563:Membrane Lipids; D009696:Nucleic Acids; D014759:Viral Envelope Proteins; C074932:prM protein, Flavivirus; C065696:glycoprotein E, Flavivirus