Use of temperature-sensitive mutants to study the morphogenesis of poliovirus.
Three temperature-sensitive (ts) mutants of poliovirus (type 1 Mahoney) were isolated after nitrous acid treatment and characterized as phenotypically RNA+. When cells were infected at 37 degrees with two of the three RNA+ ts mutants (ts109 and ts739), reduced levels of 14 S particles were synthesized. One RNA+ mutant (ts520) synthesized significant amounts of viral 14 S particle subunits. All of the mutants synthesized reduced amounts of procapsids and virions at 37 degrees. At 39.5 degrees, with all three ts mutants, the production of all virus-related particles in infected cells was markedly suppressed. Isoelectric focusing of the viral-related particles produced at 37 degrees by the ts mutants and electrophoretic analysis of their structural polypeptides revealed the following: (i) ts739 synthesized an altered VP0 polypeptide and produced 14 S particles with an altered isoelectric point; (ii) ts109 produced 14 S particles with a normal pI but containing what appeared to be an altered VP1; (iii) ts520 produced normal 14 S particles as demonstrated by their pI, the electrophoretic behavior of their constituent structural polypeptides in SDS-PAGE, their ability to self-assemble, and their ability to form procapsid-like structures when incubated in extracts from wild-type (wt) virus-infected cells. However, ts520-infected cells contained few, if any, procapsids and extracts made therefrom were unable to assemble ts520 or wt 14 S particles into detectable amounts of pI 6.8 empty capsids. These and other findings are consistent with ts739 (and probably ts109) possessing an altered structural protein and ts520 being mutant in its morphopoietic factor.
D002213:Capsid / Q000096:biosynthesis; D006367:HeLa Cells; D006801:Humans; D007526:Isoelectric Point; D009024:Morphogenesis; D009154:Mutation; D017955:Poliovirus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis*; D013696:Temperature; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis*; D015678:Viral Structural Proteins; D014771:Virion / Q000254:growth & development
D012367:RNA, Viral; D014764:Viral Proteins; D015678:Viral Structural Proteins