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. 1978 Jan;75(1):371–375. doi: 10.1073/pnas.75.1.371

Diadenosine 5',5'''-P1,P4-tetraphosphate triggers initiation of in vitro DNA replication in baby hamster kidney cells.

F Grummt
PMCID: PMC411250  PMID: 272652

Abstract

Addition of diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) to permeabilized G1-arrested baby hamster kidney cells resulted in the stimulation of DNA synthesis. No stimulation was observed in cells from exponentially growing cultures. The Ap4A-stimulated [3H]dTTP incorporation was inhibited by nalidixic acid, daunomycin, chloroquine diphosphate, EDTA, and N-ethylmaleimide. It was dose-dependent in regard to the amount of permeabilized cells and of Ap4A. Numerous replication eyes were formed in the DNA molecules of stimulated cells. Pulse-chase experiments showed that the synthesis of DNA was discontinuous, resulting in the appearance of approximately 4S Okazaki fragments and their ligation to high molecular weight DNA. These results strongly suggest that Ap4A stimulated the initiation of DNA synthesis in baby hamster kidney cells that had been arrested in G1 by serum deprival.

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Selected References

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  1. Benbow R. M., Ford C. C. Cytoplasmic control of nuclear DNA synthesis during early development of Xenopus laevis: a cell-free assay. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2437–2441. doi: 10.1073/pnas.72.6.2437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bernard O., Momparler R. L., Brent T. P. Effect of DNA polymerase on nuclei from different phases of cell cycle. Eur J Biochem. 1974 Dec 2;49(3):565–571. doi: 10.1111/j.1432-1033.1974.tb03860.x. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Francke B., Hunter T. In vitro polyoma DNA synthesis: requirement for cytoplasmic factors. J Virol. 1975 Jan;15(1):97–107. doi: 10.1128/jvi.15.1.97-107.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Friedman D. L., Mueller G. C. A nuclear system for DNA replication from synchronized HeLa cells. Biochim Biophys Acta. 1968 Jul 23;161(2):455–468. doi: 10.1016/0005-2787(68)90122-6. [DOI] [PubMed] [Google Scholar]
  6. Grummt F., Grummt I. Studies on the role of uncharged tRNA in pleiotypic response of animal cells. Eur J Biochem. 1976 Apr 15;64(1):307–312. doi: 10.1111/j.1432-1033.1976.tb10302.x. [DOI] [PubMed] [Google Scholar]
  7. Grummt F., Paul D., Grummt I. Regulation of ATP pools, rRNA and DNA synthesis in 3T3 cells in response to serum or hypoxanthine. Eur J Biochem. 1977 Jun 1;76(1):7–12. doi: 10.1111/j.1432-1033.1977.tb11564.x. [DOI] [PubMed] [Google Scholar]
  8. Grummt I., Grummt F. Control of nucleolar RNA synthesis by the intracellular pool sizes of ATP and GTP. Cell. 1976 Mar;7(3):447–453. doi: 10.1016/0092-8674(76)90175-6. [DOI] [PubMed] [Google Scholar]
  9. Grummt I., Smith V. A., Grummt F. Amino acid starvation affects the initiation frequency of nucleolar RNA polymerase. Cell. 1976 Mar;7(3):439–445. doi: 10.1016/0092-8674(76)90174-4. [DOI] [PubMed] [Google Scholar]
  10. Jazwinski S. M., Edelman G. M. Acitivity of yeast extracts in cell-free stimulation of DNA replication. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3933–3936. doi: 10.1073/pnas.73.11.3933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jazwinski S. M., Wang J. L., Edelman G. M. Initiation of replication in chromosomal DNA induced by extracts from proliferating cells. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2231–2235. doi: 10.1073/pnas.73.7.2231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson R. T., Harris H. DNA synthesis and mitosis in fused cells. II. HeLa-chick erythrocyte heterokaryons. J Cell Sci. 1969 Nov;5(3):625–643. doi: 10.1242/jcs.5.3.625. [DOI] [PubMed] [Google Scholar]
  13. Kidwell W. R., Mueller G. C. The synthesis and assembly of DNA subunits in isolated HeLa cell nuclei. Biochem Biophys Res Commun. 1969 Aug 22;36(5):756–763. doi: 10.1016/0006-291x(69)90674-3. [DOI] [PubMed] [Google Scholar]
  14. Kriegstein H. J., Hogness D. S. Mechanism of DNA replication in Drosophila chromosomes: structure of replication forks and evidence for bidirectionality. Proc Natl Acad Sci U S A. 1974 Jan;71(1):135–139. doi: 10.1073/pnas.71.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Krokan H., Wist E., Prydz H. Effect of cytosol on DNA synthesis in isolated HeLa cell nuclei. Biochem Biophys Res Commun. 1977 Mar 21;75(2):414–419. doi: 10.1016/0006-291x(77)91058-0. [DOI] [PubMed] [Google Scholar]
  16. Kumar K. V., Friedman D. L. Initiation of DNA synthesis in HeLa cell-free system. Nat New Biol. 1972 Sep 20;239(90):74–76. doi: 10.1038/newbio239074a0. [DOI] [PubMed] [Google Scholar]
  17. Murakami-Murofushi K., Mano Y. Stimulation of sea urchin DNA polymerase by protein factors. II. Formation of active complex in DNA polymerase reaction. Biochim Biophys Acta. 1977 Mar 18;475(2):254–266. doi: 10.1016/0005-2787(77)90016-8. [DOI] [PubMed] [Google Scholar]
  18. Otto B., Reichard P. Replication of polyoma DNA in isolated nuclei. V. Complementation of in vitro DNA replication. J Virol. 1975 Feb;15(2):259–267. doi: 10.1128/jvi.15.2.259-267.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Petes T. D., Newlon C. S. Structure of DNA in DNA replication mutants of yeast. Nature. 1974 Oct 18;251(5476):637–639. doi: 10.1038/251637a0. [DOI] [PubMed] [Google Scholar]
  20. Randerath K., Janeway C. M., Stephenson M. L., Zamecnik P. C. Isolation and characterization of dinucleoside tetra- and tri-phosphates formed in the presence of lysyl-sRNA synthetase. Biochem Biophys Res Commun. 1966 Jul 6;24(1):98–105. doi: 10.1016/0006-291x(66)90416-5. [DOI] [PubMed] [Google Scholar]
  21. Rao P. N., Johnson R. T. Mammalian cell fusion: studies on the regulation of DNA synthesis and mitosis. Nature. 1970 Jan 10;225(5228):159–164. doi: 10.1038/225159a0. [DOI] [PubMed] [Google Scholar]
  22. Rapaport E., Zamecnik P. C. Presence of diadenosine 5',5''' -P1, P4-tetraphosphate (Ap4A) in mamalian cells in levels varying widely with proliferative activity of the tissue: a possible positive "pleiotypic activator". Proc Natl Acad Sci U S A. 1976 Nov;73(11):3984–3988. doi: 10.1073/pnas.73.11.3984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Reiss J. R., Moffatt J. G. Dismutation reactions of nucleoside polyphosphates. 3. The synthesis of alpha, omega-dinucleoside 5'-polyphosphates. J Org Chem. 1965 Oct;30(10):3381–3387. doi: 10.1021/jo01021a029. [DOI] [PubMed] [Google Scholar]
  24. Schnös M., Inman R. B. Position of branch points in replicating lambda DNA. J Mol Biol. 1970 Jul 14;51(1):61–73. doi: 10.1016/0022-2836(70)90270-6. [DOI] [PubMed] [Google Scholar]
  25. Shimada H., Terayama H. DNA synthesis in isolated nuclei from the brains of rats at different post-partal stages and the infant rat brain cytosol factor stimulating the DNA synthesis in infant rat brain nuclei. Biochim Biophys Acta. 1972 Dec 22;287(3):415–426. doi: 10.1016/0005-2787(72)90285-7. [DOI] [PubMed] [Google Scholar]
  26. Thompson L. R., McCarthy B. J. Stimulation of nuclear DNA and RNA synthesis by cytoplasmic extracts in vitro. Biochem Biophys Res Commun. 1968 Jan 25;30(2):166–172. doi: 10.1016/0006-291x(68)90465-8. [DOI] [PubMed] [Google Scholar]
  27. Tseng B. Y., Goulian M. DNA synthesis in human lymphocyts: intermediates in DNA synthesis, in vitro and in vivo. J Mol Biol. 1975 Dec 5;99(2):317–337. doi: 10.1016/s0022-2836(75)80149-5. [DOI] [PubMed] [Google Scholar]
  28. Winnacker E. L., Magnusson G., Reichard P. Replication of polyoma DNA in isolated nuclei. I. Characterization of the system from mouse fibroblast 3T6 cells. J Mol Biol. 1972 Dec 30;72(3):523–537. doi: 10.1016/0022-2836(72)90172-6. [DOI] [PubMed] [Google Scholar]
  29. Wolstenholme D. R. Replicating DNA molecules from eggs of Drosophila melanogaster. Chromosoma. 1973 Jul 23;43(1):1–18. doi: 10.1007/BF01256731. [DOI] [PubMed] [Google Scholar]

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