Abstract
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 Cs2SO4 equilibrium centrifugation and from the finding that isotope from α-32P-labeled deoxynucleoside triphosphates was recovered in 2′(3′)-ribonucleotides after alkaline hydrolysis. Transfer occurred preferentially with [α-32P]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.
Keywords: isolated nuclei, discontinuous synthesis, deoxynucleotide starvation, hydroxyurea
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