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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Jun;72(6):2437–2441. doi: 10.1073/pnas.72.6.2437

Cytoplasmic control of nuclear DNA synthesis during early development of Xenopus laevis: a cell-free assay.

R M Benbow, C C Ford
PMCID: PMC432774  PMID: 1056038

Abstract

Nuclei isolated from nondividing cells were induced to synthesize DNA by incubation with cytoplasm from early embryos of Xenopus laevis. Numerous replication eyes were formed in the nuclear DNA molecules, and high levels of [3-H]dTTP were incorporated. With this assay a protein(s) which appears to initiate DNA synthesis was found at high levels in the cytoplasm of eggs, blastulae, or gastrulae, but only at low levels in the cytoplasm of oocytes, hatched embryos, or adult tissues.

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Selected References

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  1. Arms K. DNA synthesis by isolated embryonic nuclei of Xenopus. Dev Biol. 1971 Nov;26(3):497–502. doi: 10.1016/0012-1606(71)90079-0. [DOI] [PubMed] [Google Scholar]
  2. BEHKI R. M., SCHNEIDER W. C. Incorporation of tritiated thymidine into deoxyribonucleic acid by isolated nuclei. Biochim Biophys Acta. 1963 Jan 29;68:34–44. doi: 10.1016/0006-3002(63)90108-2. [DOI] [PubMed] [Google Scholar]
  3. Bell D. DNA synthesis in "nuclear monolayers" from BSC-1 cells infected with herpes virus. Nature. 1974 Apr 5;248(448):505–508. doi: 10.1038/248505a0. [DOI] [PubMed] [Google Scholar]
  4. Benbow R. M., Eisenberg M., Sinsheimer R. L. Multiple length DNA molecules of bacteriophage phi-X174. Nat New Biol. 1972 May 31;237(74):141–144. doi: 10.1038/newbio237141a0. [DOI] [PubMed] [Google Scholar]
  5. Benbow R. M., Mayol R. F., Picchi J. C., Sinsheimer R. L. Direction of Translation and Size of Bacteriophage phiX174 Cistrons. J Virol. 1972 Jul;10(1):99–114. doi: 10.1128/jvi.10.1.99-114.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bernard O., Brent T. P. DNA replication in isolated HeLa cell nuclei. Biochem Biophys Res Commun. 1973 Aug 21;53(4):1213–1219. doi: 10.1016/0006-291x(73)90594-9. [DOI] [PubMed] [Google Scholar]
  7. Blumenthal A. B., Kriegstein H. J., Hogness D. S. The units of DNA replication in Drosophila melanogaster chromosomes. Cold Spring Harb Symp Quant Biol. 1974;38:205–223. doi: 10.1101/sqb.1974.038.01.024. [DOI] [PubMed] [Google Scholar]
  8. Brewer E. N., Rusch H. P. DNA synthesis by isolated nuclei of Physarum polycephalum. Biochem Biophys Res Commun. 1965 Nov 8;21(3):235–241. doi: 10.1016/0006-291x(65)90277-9. [DOI] [PubMed] [Google Scholar]
  9. CHAUVEAU J., MOULE Y., ROUILLER C. Isolation of pure and unaltered liver nuclei morphology and biochemical composition. Exp Cell Res. 1956 Aug;11(2):317–321. doi: 10.1016/0014-4827(56)90107-0. [DOI] [PubMed] [Google Scholar]
  10. Callan H. G. Replication of DNA in the chromosomes of eukaryotes. Proc R Soc Lond B Biol Sci. 1972 Apr 18;181(1062):19–41. doi: 10.1098/rspb.1972.0039. [DOI] [PubMed] [Google Scholar]
  11. Case S. T., Mongeon R. L., Baker R. F. Single-stranded regions in DNA isolated from different developmental stages of the sea urchin. Biochim Biophys Acta. 1974 Apr 27;349(1):1–12. doi: 10.1016/0005-2787(74)90002-1. [DOI] [PubMed] [Google Scholar]
  12. FRIEDKIN M., WOOD H. I. V. Utilization of thymidine-C14 by bone marrow cells and isolated thymus nuclei. J Biol Chem. 1956 Jun;220(2):639–651. [PubMed] [Google Scholar]
  13. Ford C. C., Pestell R. Q., Benbow R. M. Template preferences of DNA polymerase and nuclease activities appearing during early development of Xenopus laevis. Dev Biol. 1975 Mar;43(1):175–188. doi: 10.1016/0012-1606(75)90139-6. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Friedman D. L. On the mechanism of DNA replication in isolated nuclei from HeLa cells. Biochim Biophys Acta. 1974 Jul 24;353(4):447–462. doi: 10.1016/0005-2787(74)90051-3. [DOI] [PubMed] [Google Scholar]
  16. Gurdon J. B., Laskey R. A. The transplantation of nuclei from single cultured cells into enucleate frogs' eggs. J Embryol Exp Morphol. 1970 Sep;24(2):227–248. [PubMed] [Google Scholar]
  17. Gurdon J. B. On the origin and persistence of a cytoplasmic state inducing nuclear DNA synthesis in frogs' eggs. Proc Natl Acad Sci U S A. 1967 Aug;58(2):545–552. doi: 10.1073/pnas.58.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gurdon J. B., Woodland H. R. The cytoplasmic control of nuclear activity in animal development. Biol Rev Camb Philos Soc. 1968 May;43(2):233–267. doi: 10.1111/j.1469-185x.1968.tb00960.x. [DOI] [PubMed] [Google Scholar]
  19. Hershey H. V., Stieber J. F., Mueller G. C. Dna synthesis in isolated HeLa nuclei. A system for continuation of replication in vivo. Eur J Biochem. 1973 Apr;34(2):383–394. doi: 10.1111/j.1432-1033.1973.tb02770.x. [DOI] [PubMed] [Google Scholar]
  20. Inman R. B., Schnös M. Structure of branch points in replicating DNA: presence of single-stranded connections in lambda DNA branch points. J Mol Biol. 1971 Mar 14;56(2):319–325. doi: 10.1016/0022-2836(71)90467-0. [DOI] [PubMed] [Google Scholar]
  21. Kasamatsu H., Grossman L. I., Robberson D. L., Watson R., Vinograd J. The replication and structure of mitochondrial DNA in animal cells. Cold Spring Harb Symp Quant Biol. 1974;38:281–288. doi: 10.1101/sqb.1974.038.01.031. [DOI] [PubMed] [Google Scholar]
  22. Kemper B. W., Pratt W. B., Aronow L. Nucleic acid synthesis in intact nuclei isolated from mouse fibroblasts. Characterization of the system and effects of glucocorticoids. Mol Pharmacol. 1969 Sep;5(5):507–531. [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Lagunoff D. Thymidine-5'-triphosphate incorporation into DNA by rat thymus nuclei. Exp Cell Res. 1969 Apr;55(1):53–56. doi: 10.1016/0014-4827(69)90454-6. [DOI] [PubMed] [Google Scholar]
  26. Lynch W. E., Brown R. F., Umeda T., Langreth S. G., Lieberman I. Synthesis of deoxyribonucleic acid by isolated liver nuclei. J Biol Chem. 1970 Aug 10;245(15):3911–3916. [PubMed] [Google Scholar]
  27. MAZIA D., HINEGARDNER R. T. Enzymes of DNA synthesis in nuclei of sea urchin embryos. Proc Natl Acad Sci U S A. 1963 Jul;50:148–156. doi: 10.1073/pnas.50.1.148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. 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]
  31. Waqar M. A., Burgoyne L. A., Atkinson M. R. Deoxyribonucleic acid synthesis in mammalian nuclei. Incorporation of deoxyribonucleotides and chain-terminating nucleotide analogues. Biochem J. 1971 Mar;121(5):803–809. doi: 10.1042/bj1210803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wilson D. A., Thomas C. A., Jr Palindromes in chromosomes. J Mol Biol. 1974 Mar 25;84(1):115–138. doi: 10.1016/0022-2836(74)90216-2. [DOI] [PubMed] [Google Scholar]
  33. Winnacker E. L., Magnusson G., Reichard P. Synthesis of polyoma DNA by isolated nuclei. Biochem Biophys Res Commun. 1971 Aug 20;44(4):952–957. doi: 10.1016/0006-291x(71)90804-7. [DOI] [PubMed] [Google Scholar]
  34. 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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