title: Signal peptide peptidase-catalyzed cleavage of hepatitis C virus core protein is dispensable for virus budding but destabilizes the viral capsid.
abstract: The capsid of hepatitis C virus (HCV) particles is considered to be composed of the mature form (p21) of core protein. Maturation to p21 involves cleavage of the transmembrane domain of the precursor form (p23) of core protein by signal peptide peptidase (SPP), a cellular protease embedded in the endoplasmic reticulum membrane. Here we have addressed whether SPP-catalyzed maturation to p21 is a prerequisite for HCV particle morphogenesis in the endoplasmic reticulum. HCV structural proteins were expressed by using recombinant Semliki Forest virus replicon in mammalian cells or recombinant baculovirus in insect cells, because these systems have been shown to allow the visualization of HCV budding events and the isolation of HCV-like particles, respectively. Inhibition of SPP-catalyzed cleavage of core protein by either an SPP inhibitor or HCV core mutations not only did not prevent but instead tended to facilitate the observation of viral buds and the recovery of virus-like particles. Remarkably, although maturation to p21 was only partially inhibited by mutations in insect cells, p23 was the only form of core protein found in HCV-like particles. Finally, newly developed assays demonstrated that p23 capsids are more stable than p21 capsids. These results show that SPP-catalyzed cleavage of core protein is dispensable for HCV budding but decreases the stability of the viral capsid. We propose a model in which p23 is the form of HCV core protein committed to virus assembly, and cleavage by SPP occurs during and/or after virus budding to predispose the capsid to subsequent disassembly in a new cell.
authors.affiliation: INSERM, Equipe AVENIR, Institut Cochin, 75014 Paris, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Vauloup-Fellous
authors.forename: Véronique
authors.initials: V
authors.lastname: Pène
authors.forename: Julie
authors.initials: J
authors.lastname: Garaud-Aunis
authors.forename: Francis
authors.initials: F
authors.lastname: Harper
authors.forename: Sabine
authors.initials: S
authors.lastname: Bardin
authors.forename: Yannick
authors.initials: Y
authors.lastname: Suire
authors.forename: Evelyne
authors.initials: E
authors.lastname: Pichard
authors.forename: Alain
authors.initials: A
authors.lastname: Schmitt
authors.forename: Philippe
authors.initials: P
authors.lastname: Sogni
authors.forename: Gérard
authors.initials: G
authors.lastname: Pierron
authors.forename: Pascale
authors.initials: P
authors.lastname: Briand
authors.forename: Arielle R
authors.initials: AR
authors.lastname: Rosenberg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000502:physiology*; D002213:Capsid / Q000378:metabolism*; D002384:Catalysis; D002478:Cells, Cultured; D016174:Hepacivirus / Q000502:physiology* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009024:Morphogenesis; D018411:Spodoptera; D014758:Viral Core Proteins / Q000378:metabolism*; D014771:Virion / Q000502:physiology; D019065:Virus Assembly
chemical_list: D014758:Viral Core Proteins; D016282:Aspartic Acid Endopeptidases; C460085:signal peptide peptidase