Extensive symmetrical transcription of Simian Virus 40 DNA in virus-yielding cells.
Rapidly labeled RNA was extracted from monkey cells after infection with Simian Virus 40 (SV40) and exposure to short pulses of [5-(3)H]uridine late in infection. When this RNA was self-annealed, it became resistant to digestion with ribonuclease. The fraction of RNA that resisted the ribonuclease treatment decreased with increased labeling time, or when a short pulse of radioactivity was followed by incubation with unlabeled uridine and actinomycin D. The RNase-resistant RNA was isolated by chromatography on Sephadex G-100 and shown to be double-stranded by its susceptibility to ribonuclease as a function of salt concentration and temperature. This behavior was not due to RNA-DNA hybrid formation, since deoxyribonuclease had no effect upon the double-stranded molecules, even after their denaturation. The relation of the double-stranded RNA to SV40 was demonstrated by the hybridization of about 50% (corrected value, >90%) of the separated RNA strands with component I of SV40 DNA from plaque-purified virus. After self-annealing in formamide at low temperature, about 10% of the rapidly labeled, viral RNA sedimented at 13 S. This value corresponds in size to about 60% of the SV40 DNA.These observations indicate that late in infection of monkey cells, SV40 DNA is transcribed symmetrically over a considerable portion of its length, and that subsequently some sequences from one or both of the RNA strands are degraded.
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004247:DNA; D004279:DNA, Viral / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology; D003851:Deoxyribonucleases; D005815:Genetic Code; D000882:Haplorhini; D007553:Isotope Labeling; D007668:Kidney / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009994:Osmolar Concentration; D012367:RNA, Viral / Q000096:biosynthesis*; D012260:Ribonucleases; D013539:Simian virus 40; D013696:Temperature; D013997:Time Factors; D014316:Tritium; D014529:Uridine / Q000378:metabolism
D004279:DNA, Viral; D012367:RNA, Viral; D014316:Tritium; D003609:Dactinomycin; D004247:DNA; D003851:Deoxyribonucleases; D012260:Ribonucleases; D014529:Uridine