Replication of polyoma DNA in isolated nuclei. V. Complementation of in vitro DNA replication.
AUTOR(ES)
Otto, B
RESUMO
Nuclei from polyoma-infected 3T6 fibroblasts elongate in vitro the progeny strands of the replicative intermediates of polyoma DNA. When high concentrations of such nuclei were incubated, short DNA fragments were formed and subsequently added onto growing progeny strands. When nuclei were repeatedly washed with buffer containing detergent and then incubated at low concentrations. DNA synthesis was decreased. In particular, the joining process was reduced, resulting in an accumulation of short DNA fragments. All aspects of the synthetic capacity of the nuclei were restored by addition of cytoplasmic extract. Additions of purified enzymes (polynucleotide ligase from calf thymus or Escherichia coli together with E. coli DNA polymerase I) increased the joining function of the nuclei. The system can be used for the identification of the enzymatic steps concerned with polyoma DNA replication.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=354448Documentos Relacionados
- Strandedness of newly synthesized short pieces of polyoma DNA from isolated nuclei.
- DNA synthesis in polyoma virus infection. V. Kinetic evidence for two requirements for protein synthesis during viral DNA replication.
- Replication of polyoma DNA in isolated nuclei: analysis of replication fork movement.
- Polyoma DNA synthesis in isolated nuclei: evidence for defective replication forks.
- Species-specific in vitro synthesis of DNA containing the polyoma virus origin of replication.