title: Acid-activated structural reorganization of the Rift Valley fever virus Gc fusion protein.
abstract: The entry of the enveloped Rift Valley fever virus (RVFV) into its host cell is mediated by the viral glycoproteins Gn and Gc. We investigated the RVFV entry process and, in particular, its pH-dependent activation mechanism using our recently developed nonspreading-RVFV-particle system. Entry of the virus into the host cell was efficiently inhibited by lysosomotropic agents that prevent endosomal acidification and by compounds that interfere with dynamin- and clathrin-dependent endocytosis. Exposure of plasma membrane-bound virions to an acidic pH (<pH 6) equivalent to the pH of late endolysosomal compartments allowed the virus to bypass the endosomal route of infection. Acid exposure of virions in the absence of target membranes triggered the class II-like Gc fusion protein to form extremely stable oligomers that were resistant to SDS and temperature dissociation and concomitantly compromised virus infectivity. By targeted mutagenesis of conserved histidines in Gn and Gc, we demonstrated that mutation of a single histidine (H857) in Gc completely abrogated virus entry, as well as acid-induced Gc oligomerization. In conclusion, our data suggest that after endocytic uptake, RVFV traffics to the acidic late endolysosomal compartments, where histidine protonation drives the reorganization of the Gc fusion protein that leads to membrane fusion.
authors.affiliation: Department of Infectious Diseases and Immunology, Virology Division, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: S M
authors.initials: SM
authors.lastname: de Boer
authors.forename: J
authors.initials: J
authors.lastname: Kortekaas
authors.forename: L
authors.initials: L
authors.lastname: Spel
authors.forename: P J M
authors.initials: PJ
authors.lastname: Rottier
authors.forename: R J M
authors.initials: RJ
authors.lastname: Moormann
authors.forename: B J
authors.initials: BJ
authors.lastname: Bosch
mesh_terms: D000143:Acids / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D017931:DNA Primers; D004330:Drosophila; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005434:Flow Cytometry; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence; D011487:Protein Conformation; D012296:Rift Valley fever virus / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000143:Acids; D017931:DNA Primers; D014760:Viral Fusion Proteins