Coinfection with recombinant vaccinia viruses expressing poliovirus P1 and P3 proteins results in polyprotein processing and formation of empty capsid structures.
The assembly process of poliovirus occurs via an ordered proteolytic processing of the capsid precursor protein, P1, by the virus-encoded proteinase 3CD. To further delineate this process, we have isolated a recombinant vaccinia virus which expresses, upon infection, the poliovirus P1 capsid precursor polyprotein with an authentic carboxy terminus. Coinfection of HeLa cells with the P1-expressing vaccinia virus and with a second recombinant vaccinia virus which expresses the poliovirus proteinase 3CD resulted in the correct processing of P1 to yield the three individual capsid proteins VP0, VP3, and VP1. When extracts from coinfected cells were fractionated on sucrose density gradients, the VP0, VP3, and VP1 capsid proteins were immunoprecipitated with type 1 poliovirus antisera from fractions corresponding to a sedimentation consistent for poliovirus 75S procapsids. Examination of these fractions by electron microscopy revealed structures which lacked electron-dense cores and which corresponded in size and shape to those expected for poliovirus empty capsids. We conclude that the expression of the two poliovirus proteins P1 and 3CD in coinfected cells is sufficient for the correct processing of the capsid precursor to VP0, VP3, and VP1 as well as for the assembly of poliovirus empty capsid-like structures.
Ansardi
D C
DC
Department of Microbiology, University of Alabama, Birmingham, 35294.
Porter
D C
DC
Morrow
C D
CD
D000086042:3C Viral Proteases; D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics*; D036022:Capsid Proteins; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry; D015870:Gene Expression; D005814:Genes, Viral; D006367:HeLa Cells / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D011051:Poliomyelitis / Q000235:genetics; D017955:Poliovirus / Q000201:enzymology / Q000235:genetics* / Q000648:ultrastructure; D011499:Protein Processing, Post-Translational; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins
D036022:Capsid Proteins; D004279:DNA, Viral; C442540:VP1 protein, Poliovirus; C442542:VP3 protein, Poliovirus; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases