RNA-linked short DNA fragments during polyoma replication.
During in vitro incubation, nuclei from polyoma-infected cells elongate the daughter strands of the replicative intermediate of polyoma DNA. This process is now shown to involve the transient formation of short fragments (4-5 S), a process that is stimulated by the addition of ribonucleoside triphosphates. The presence of stretches of RNA at the 5'-end of short DNA chains was determined from Cs(2)SO(4) equilibrium centrifugation and from the finding that isotope from alpha-(32)P-labeled deoxynucleoside triphosphates was recovered in 2'(3')-ribonucleotides after alkaline hydrolysis. Transfer occurred preferentially with [alpha-(32)P]dCTP as substrate. Starvation for deoxynucleotides by in vivo treatment with hydroxyurea resulted in the accumulation of short fragments that are deficient in RNA. Our results suggest that a late step during the discontinuous synthesis of polyoma DNA is selectively inhibited when deoxynucleotides are in short supply.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002247:Carbon Isotopes; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000382:microbiology; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D003597:Cytosine Nucleotides / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000032:analysis; D006918:Hydroxyurea / Q000494:pharmacology; D051379:Mice; D009691:Nucleic Acid Denaturation; D010759:Phosphorus Isotopes; D011120:Polyomavirus / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000378:metabolism; D014316:Tritium; D014500:Uracil Nucleotides / Q000378:metabolism; D014779:Virus Replication
D002247:Carbon Isotopes; D003597:Cytosine Nucleotides; D004279:DNA, Viral; D010759:Phosphorus Isotopes; D012367:RNA, Viral; D013942:Thymine Nucleotides; D014500:Uracil Nucleotides; D014316:Tritium; D000255:Adenosine Triphosphate; D006918:Hydroxyurea