Mutations in VP1 of poliovirus specifically affect both encapsidation and release of viral RNA.
The phenotypic defects of two type 1 Mahoney poliovirus mutants, termed VP1-101 and VP1-102, were caused by two different small deletions in the region of the RNA genome encoding the amino terminus of the capsid protein VP1. This portion of VP1 was unresolved in the three-dimensional structure of the poliovirion, buried within the virion, and likely to interact with the viral RNA. Both VP1-101 and VP1-102 showed a diminished ability to enter CV1 but not HeLa cells; both mutants formed plaques on CV1 and HeLa cells that were smaller than wild type. Neither the rate of binding to cells nor the rate of subsequent receptor-dependent conformational change of the mutant poliovirions was affected. However, both mutants displayed delayed kinetics of RNA release compared with wild-type virus. One of the mutants, VP1-102, also displayed a defect in viral morphogenesis: 75S empty capsids formed normally, but 150S particles that contained RNA accumulated much more slowly. We suggest that the VP1-102 mutation affects RNA encapsidation as well as RNA release, whereas the VP1-101 mutation affects only RNA release. Therefore, RNA packaging and RNA release are genetically linked but can be mutated separately in different VP1 alleles, and both processes involve the amino terminus of VP1.
Kirkegaard
K
K
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309.
D000818:Animals; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D002460:Cell Line; D002872:Chromosome Deletion; D005814:Genes, Viral; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008961:Models, Structural; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D017955:Poliovirus / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection; D010948:Viral Plaque Assay; D015678:Viral Structural Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D012367:RNA, Viral; C442540:VP1 protein, Poliovirus; D015678:Viral Structural Proteins