Viral RNAs associated with ribosomes in Sindbis virus-infected HeLa cells.
Virus specific RNA ribosome complexes were isolated by sucrose density gradient centrifugation of cytoplasmic extracts from HeLa cells infected at 42 C with an RNA(+) mutant (ts2) of Sindbis virus. Viral RNA-ribosome complexes were accumulated by infected cells treated with sodium fluoride and cycloheximide. The RNA-ribosome complexes were characterized by (i) their sensitivity to the action of ribonuclease or ethylenediaminetetraacetic acid, (ii) their density in cesium chloride gradients, and (iii) presence of host ribosomes and viral RNAs. The viral RNAs were isolated and characterized. The results showed that two species of single-stranded RNAs (a 28s and 18 to 15s species) were associated with the complexes. Base composition analysis of the viral RNAs indicated that both species had a higher adenine content than the 42s or 26s forms of viral RNAs. The RNAs associated with the ribosome complexes were virus specific since they annealed with denatured double-stranded RNAs from the infected cells. Little or no 42S RNA was associated with the RNA-ribosome complexes. The results suggest that the 28s and 18 to 15s forms of RNAs may represent viral messenger RNAs.
D000225:Adenine / Q000032:analysis; D002247:Carbon Isotopes; D002499:Centrifugation, Density Gradient; D003513:Cycloheximide / Q000494:pharmacology; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D005459:Fluorides / Q000494:pharmacology; D006367:HeLa Cells / Q000187:drug effects / Q000382:microbiology*; D006801:Humans; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010759:Phosphorus Isotopes; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D012260:Ribonucleases; D012270:Ribosomes; D012845:Sindbis Virus / Q000254:growth & development / Q000302:isolation & purification; D014316:Tritium
D002247:Carbon Isotopes; D010759:Phosphorus Isotopes; D012333:RNA, Messenger; D012367:RNA, Viral; D014316:Tritium; D012313:RNA; D003513:Cycloheximide; D004492:Edetic Acid; D012260:Ribonucleases; D000225:Adenine; D005459:Fluorides