Isopycnic separation of subcellular components from poliovirus-infected and normal HeLa cells.
A rapid method for determining the density of ribonucleoprotein particles and complexes has been developed. The method involves glutaraldehyde fixation of sucrose gradient fractions and immediate centrifugation for 5 hours through preformed cesium chloride gradients. There is little or no aggregation of particles, so that components which co-sediment in sucrose gradients are resolved by the cesium chloride gradient. By this method the densities of HeLa cell ribosomes, polyribosomes, and subribosomal particles have been determined. Furthermore, the poliovirus replication complex has been separated from polyribosomes and its density has been found to be unaffected by treatment with ethylene-diaminetetraacetic acid.
D000447:Aldehydes; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002712:Chlorides; D003588:Cytopathogenic Effect, Viral; D006367:HeLa Cells / Q000166:cytology* / Q000382:microbiology*; D008722:Methods; D017955:Poliovirus / Q000472:pathogenicity*; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis; D012270:Ribosomes / Q000032:analysis*; D014529:Uridine / Q000378:metabolism
D000447:Aldehydes; D002712:Chlorides; D012367:RNA, Viral; D002586:Cesium; D014529:Uridine