Isolation of a viral polypeptide associated with poliovirus RNA polymerase.
Poliovirus-infected HeLa cells were labeled with radioactive methionine or phenylalanine and subjected to a new purification procedure for the viral induced RNA polymerase activity. Detergent-solubilized polymerase activity was purified by precipitation with 2 M LiCl and sedimentation through sucrose gradients. Approximately 0.001% of the incorporated amino acid radio-activity sediments with the peak of polymerase activity. Gradient fractions comprising the polymerase activity peak were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and found to contain predominantly one virus-specific polypeptide. Polyacrylamide gel electrophoresis also reveals that this purified polypeptide migrates with a 58,000 molecular weight noncapsid polio-virus polypeptide.
D002499:Centrifugation, Density Gradient; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D010455:Peptides / Q000302:isolation & purification*; D010649:Phenylalanine / Q000378:metabolism; D017955:Poliovirus / Q000201:enzymology*; D013462:Sulfur Radioisotopes; D014316:Tritium
D010455:Peptides; D013462:Sulfur Radioisotopes; D014316:Tritium; D010649:Phenylalanine; D008715:Methionine; D012321:DNA-Directed RNA Polymerases