Protein synthesis directed by an RNA temperature-sensitive mutant of Sindbis virus.
The structural proteins of wild-type Sindbis virus were shown to arise by posttranslational cleavage of larger precursors. The proteins synthesized in wildtype infection were compared with those specified by ts-11, a temperature-sensitive mutant unable to synthesize viral RNA at the restrictive temperature. Abnormally large, virus-specific proteins were found in the mutant-infected cells after the shift from 28 C to 41.5 C. These large polypeptides were presumably precursors which were cleaved too rapidly to be detected in the wild-type infection. The largest had a molecular weight of 133,000 and was the same size as the apparent precursor detected during infection with a group of Sindbis mutants which could not form nucleocapsids at the nonpermissive temperature. The stability of ts-11-specific RNA synthesis, after shift from permissive to restrictive conditions, differed from that in cells infected by wild-type virus, indicating that the virus had a genetic lesion in an enzyme involved in RNA synthesis. This mutation might have caused the precursor to fold incorrectly so that it could not be cleaved. The possibility cannot be excluded, however, that a second lesion in an uncharacterized viral function, such as a protease, was the cause of the accumulation of the precursors.
D000818:Animals; D002247:Carbon Isotopes; D002499:Centrifugation, Density Gradient; D002642:Chick Embryo; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D009154:Mutation; D010455:Peptides / Q000032:analysis; D011498:Protein Precursors; D012367:RNA, Viral / Q000096:biosynthesis*; D012845:Sindbis Virus / Q000032:analysis / Q000254:growth & development / Q000378:metabolism*; D013696:Temperature; D014316:Tritium; D014529:Uridine / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis*
D002247:Carbon Isotopes; D010455:Peptides; D011498:Protein Precursors; D012367:RNA, Viral; D014764:Viral Proteins; D014316:Tritium; D007930:Leucine; D014529:Uridine