title: PE2 cleavage mutants of Sindbis virus: correlation between viral infectivity and pH-dependent membrane fusion activation of the spike heterodimer.
abstract: The spike glycoprotein E2 of Sindbis virus (SIN) is synthesized in the infected cell as a PE2 precursor protein, which matures through cleavage by a cellular furin-like protease. Previous work has shown that SIN mutants impaired in PE2 cleavage are noninfectious on BHK-21 cells, the block in infection being localized at a step after virus-receptor interaction but prior to RNA replication. Here, we studied the membrane fusion properties of SIN PE2 cleavage mutants and observed that these viruses are impaired in their ability to form an E1 homotrimer and to fuse with liposomes at a mildly acidic pH. The block in spike rearrangement and fusion could be overridden by exposure of the mutant viruses to very low pH (<4.5). Cleavage mutants with second-site resuscitating mutations in PE2 were highly infectious for BHK-21 cells. The ability of these viruses to form E1 homotrimers and to fuse at a mildly acidic pH was completely restored despite a sustained lack of PE2 cleavage.
authors.affiliation: Molecular Virology Section, Department of Medical Microbiology, University of Groningen, 9713 AV Groningen, The Netherlands.
authors.forename: J M
authors.initials: JM
authors.lastname: Smit
authors.forename: W B
authors.initials: WB
authors.lastname: Klimstra
authors.forename: K D
authors.initials: KD
authors.lastname: Ryman
authors.forename: R
authors.initials: R
authors.lastname: Bittman
authors.forename: R E
authors.initials: RE
authors.lastname: Johnston
authors.forename: J
authors.initials: J
authors.lastname: Wilschut
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D019281:Dimerization; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000737:chemistry*; D009154:Mutation; D011498:Protein Precursors / Q000737:chemistry*; D012845:Sindbis Virus / Q000472:pathogenicity*; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008562:Membrane Glycoproteins; D011498:Protein Precursors; D014759:Viral Envelope Proteins; C022012:glycoprotein E1, Sindbis virus; C064950:glycoprotein E2, Sindbis virus