Isolation and properties of an RNA polymerase from influenza virus-infected cells.
Structures with RNA polymerase activity were isolated from influenza virus-infected cells, and consisted of ribonucleoprotein (RNP) complexes, similar in morphology to the viral internal component or nucleocapsid. The isolation procedure involved fractionation of infected cells in a discontinuous sucrose gradient, in which enzyme activity was concentrated in a fraction of intermediate density which contains both smooth and rough cytoplasmic membranes. The RNPs with polymerase activity were further purified in a velocity gradient, after which the peak fractions showed a 35-fold purification of the polymerase activity when compared with cytoplasmic extracts. The NP polypeptide, which is the subunit of the virion RNP, was the only virus-specific polypeptide detected in these RNP structures.
D000818:Animals; D002247:Carbon Isotopes; D002417:Cattle; D002458:Cell Fractionation; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D003593:Cytoplasm / Q000201:enzymology; D012321:DNA-Directed RNA Polymerases / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005944:Glucosamine / Q000378:metabolism; D007668:Kidney; D008854:Microscopy, Electron; D009698:Nucleoproteins / Q000032:analysis; D009711:Nucleotides / Q000378:metabolism; D009975:Orthomyxoviridae / Q000032:analysis; D010455:Peptides / Q000032:analysis; D011492:Protein Hydrolysates / Q000378:metabolism; D013194:Staining and Labeling; D014316:Tritium; D014764:Viral Proteins / Q000032:analysis
D002247:Carbon Isotopes; D009698:Nucleoproteins; D009711:Nucleotides; D010455:Peptides; D011492:Protein Hydrolysates; D014764:Viral Proteins; D014316:Tritium; D012321:DNA-Directed RNA Polymerases; D005944:Glucosamine