Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1978 Jul;5(7):2535–2545. doi: 10.1093/nar/5.7.2535

Okazaki pieces grow opposite to the replication fork direction during simian virus 40 DNA replication.

G Kaufmann, R Bar-Shavit, M L DePamphilis
PMCID: PMC342183  PMID: 209411

Abstract

Simian virus 40 replicating DNA was pulse labeled with alpha-32P-dATP using an acellular DNA replication system. Nascent DNA chains of less than 200 nucleotides (Okazaki pieces) were then isolated from the denatured replicating DNA by electrosieving through a polyacrylamide gel column. The purified Okazaki pieces were hybridized to separated strands of Bg1(1)+Hpa1 simian virus 40 DNA restriction fragments immobilized on nitrocellulose filters. Only strands with polarity of the DNA replication fork direction hybridized with Okazaki pieces. Hence, Okazaki pieces in simian virus 40 are synthesized against the DNA replication fork direction.

Full text

PDF
2536

Images in this article

2536Image
on p.2536
2538Image
on p.2538
2541Image
on p.2541
2543Image
on p.2543

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aloni Y., Dhar R., Laub O., Horowitz M., Khoury G. Novel mechanism for RNA maturation: the leader sequences of simian virus 40 mRNA are not transcribed adjacent to the coding sequences. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3686–3690. doi: 10.1073/pnas.74.9.3686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. DePamphilis M. L., Berg P. Requirement of a Cytoplasmic Fraction for Synthesis of SV40 Deoxyribonucleic Acid in Isolated Nuclei*. J Biol Chem. 1975 Jun 10;250(11):4348–4354. [PubMed] [Google Scholar]
  3. Fareed G. C., Salzman N. P. Intermediate in SV40 DNA chain growth. Nat New Biol. 1972 Aug 30;238(87):274–277. doi: 10.1038/newbio238274a0. [DOI] [PubMed] [Google Scholar]
  4. Flory P. J., Jr Strandedness of newly synthesized short pieces of polyoma DNA from isolated nuclei. Nucleic Acids Res. 1977;4(5):1449–1464. doi: 10.1093/nar/4.5.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hunter T., Francke B., Bacheler L. In vitro polyoma DNA synthesis: asymmetry of short DNA chains. Cell. 1977 Dec;12(4):1021–1028. doi: 10.1016/0092-8674(77)90166-0. [DOI] [PubMed] [Google Scholar]
  6. Kaufmann G., Anderson S., DePamphilis M. L. RNA primers in Simian virus 40 DNA replication. II. Distribution of 5' terminal oligoribonucleotides in nascent DNA. J Mol Biol. 1977 Nov 5;116(3):549–567. doi: 10.1016/0022-2836(77)90083-3. [DOI] [PubMed] [Google Scholar]
  7. Kelly T. J., Jr, Nathans D. The genome of simian virus 40. Adv Virus Res. 1977;21:85–173. doi: 10.1016/s0065-3527(08)60762-9. [DOI] [PubMed] [Google Scholar]
  8. Perlman D., Huberman J. A. Asymmetric Okazaki piece synthesis during replication of simian virus 40 DNA in vivo. Cell. 1977 Dec;12(4):1029–1043. doi: 10.1016/0092-8674(77)90167-2. [DOI] [PubMed] [Google Scholar]
  9. Pigiet V., Eliasson R., Reichard P. Replication of polyoma DNA in isolated nuclei. 3. The nucleotide sequence at the RNA-DNA junction of nascent strands. J Mol Biol. 1974 Mar 25;84(1):197–216. doi: 10.1016/0022-2836(74)90222-8. [DOI] [PubMed] [Google Scholar]
  10. Reichard P., Eliasson R., Söderman G. Initiator RNA in discontinuous polyoma DNA synthesis. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4901–4905. doi: 10.1073/pnas.71.12.4901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Seale R. L. Studies on the mode of segregation of histone nu bodies during replication in HeLa cells. Cell. 1976 Nov;9(3):423–429. doi: 10.1016/0092-8674(76)90087-8. [DOI] [PubMed] [Google Scholar]
  12. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  13. Tye B. K., Lehman I. R. Excision repair of uracil incorporated in DNA as a result of a defect in dUTPase. J Mol Biol. 1977 Dec 5;117(2):293–306. doi: 10.1016/0022-2836(77)90128-0. [DOI] [PubMed] [Google Scholar]
  14. Weintraub H. Cooperative alignment of nu bodies during chromosome replication in the presence of cycloheximide. Cell. 1976 Nov;9(3):419–422. doi: 10.1016/0092-8674(76)90086-6. [DOI] [PubMed] [Google Scholar]
  15. Westphal H. SV40 DNA strand selection by Escherichia coli RNA polymerase. J Mol Biol. 1970 Jun 14;50(2):407–420. doi: 10.1016/0022-2836(70)90201-9. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES