Poliovirus mutants at histidine 195 of VP2 do not cleave VP0 into VP2 and VP4.
The final stage of poliovirus assembly is characterized by a cleavage of the capsid precursor protein VP0 into VP2 and VP4. This cleavage is thought to be autocatalytic and dependent on RNA encapsidation. Analysis of the poliovirus empty capsid structure has led to a mechanistic model for VP0 cleavage involving a conserved histidine residue that is present in the surrounding environment of the VP0 cleavage site. Histidine 195 of VP2 (2195H) is hypothesized to activate local water molecules, thus initiating a nucleophilic attack at the scissile bond. To test this hypothesis, 2195H mutants were constructed and their phenotypes were characterized. Consistent with the requirement of VP0 cleavage for poliovirus infectivity, all 2195H mutants were nonviable upon introduction of the mutant genomes into HeLa cells. Replacement of 2195H with threonine or arginine resulted in the assembly of a highly unstable 150S virus particle. Further analyses showed that these particles contain genomic RNA and uncleaved VP0, criteria associated with the provirion assembly intermediate. These data support the involvement of 2195H in mediating VP0 cleavage during the final stages of virus assembly.
Hindiyeh
M
M
Department of Microbiology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
Li
Q H
QH
Basavappa
R
R
Hogle
J M
JM
Chow
M
M
D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002499:Centrifugation, Density Gradient; D006367:HeLa Cells; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D017955:Poliovirus / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D011498:Protein Precursors / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014771:Virion; D019065:Virus Assembly; D014779:Virus Replication
D036022:Capsid Proteins; D011498:Protein Precursors; C442539:VP0 protein, Poliovirus; C442541:VP2 protein, Poliovirus; C060339:VP4 protein, Rotavirus; D006639:Histidine