title: Processing of proteinase precursors and their effect on hepatitis A virus particle formation.
abstract: Proteolytic processing of the picornaviral polyprotein mediated by the differential action of virus-encoded proteinase(s) is pivotal to both RNA genome replication and capsid formation. Possibly to enlarge the array of viral proteins, picornaviral polyprotein processing results in intermediate and mature products which apparently have distinct functions within the viral life cycle. For hepatitis A virus (HAV), we report here on the autoproteolysis of precursor polypeptides comprising the only viral proteinase, 3Cpro, and on their role in viral particle formation. Following transient expression of a nested set of 3Cpro-containing proteins (P3, 3ABC, 3BCD, 3CD, 3BC, and 3C) in eukaryotic cells, the extent of processing was determined by analyzing the cleavage products. The 3C/3D site was more efficiently cleaved than those at the 3A/3B and 3B/3C sites, leading to the accumulation of the intermediate product 3ABC. In the absence of 3A from the precursor, cleavage at the 3B/3C site was further reduced and a switch to an alternative 3C/3D site was observed. Coexpression of various parts of P3 with the precursor of the viral structural proteins P1-2A showed that all 3C-containing intermediates cleaved P1-2A with almost equal efficiency; however, viral particles carrying the neutralizing epitope form much more readily in the presence of the complete P3 domain than with parts of it. These data support the notion that efficient liberation of structural proteins from P1-2A is necessary but not sufficient for productive HAV capsid formation and suggest that the polypeptides flanking 3Cpro promote the assembly of viral particles.
authors.affiliation: Institute for Medical Microbiology, Medical University of Lübeck, 23538 Lübeck, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Probst
authors.forename: M
authors.initials: M
authors.lastname: Jecht
authors.forename: V
authors.initials: V
authors.lastname: Gauss-Müller
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D010450:Endopeptidases / Q000378:metabolism*; D004792:Enzyme Precursors / Q000378:metabolism*; D006507:Hepatovirus / Q000201:enzymology / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D014771:Virion
chemical_list: D017931:DNA Primers; D004792:Enzyme Precursors; D010450:Endopeptidases