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. 1982 Oct 11;10(19):6191–6205. doi: 10.1093/nar/10.19.6191

In the higher plant Pisum sativum maturation of nascent DNA is blocked by cycloheximide, but only after 4-8 replicons are joined.

J B Schvartzman, J Van't Hof
PMCID: PMC320960  PMID: 7145718

Abstract

Velocity sedimentation in alkaline sucrose gradients, single cell autoradiography and cytophotometry were used to determine if protein synthesis is required for the maturation of nascent replicons to chromosomal-sized molecules in cultured pea-root cells. The results obtained showed that cycloheximide at 5 and 10 microgram/ml, added either before or during labeling with tritiated thymidine, blocked maturation of nascent DNA at an intermediate size of 72-140 X 10(6) daltons single-stranded DNA. To reach this size, nascent replicons - which are 18 X 10(6) daltons single-stranded DNA each - were replicated and groups of 4-8 replicons were joined even though protein synthesis was reduced to 15% of the control. Further maturation of the nascent molecules to chromosomal size, however, was prevented and this resulted in the accumulation of nascent molecules in the 72-140 X 10(6) daltons range. The experiments also showed that the joining of nascent replicons is not an absolute function of late S or G2 phase of the cell cycle, since cells treated with cycloheximide and blocked in mid-S phase had nascent DNA of a size corresponding to 4-8 joined replicons. Finally, the results support the hypothesis that at least one step in the process of nascent DNA maturation may require replication, during late-S phase, of DNA segments that are interspersed within replicon-clusters that replicate early in the S phase.

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Selected References

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

  1. Bloch D. P., Macquigg R. A., Brack S. D., Wu J. R. The syntheses of deoxyribonucleic acid and histone in the onion root meristem. J Cell Biol. 1967 Jun;33(3):451–467. doi: 10.1083/jcb.33.3.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carrier W. L., Setlow R. B. Paper strip method for assaying gradient fractions containing radioactive macromolecules. Anal Biochem. 1971 Oct;43(2):427–432. doi: 10.1016/0003-2697(71)90272-7. [DOI] [PubMed] [Google Scholar]
  3. Edenberg H. J., Huberman J. A. Eukaryotic chromosome replication. Annu Rev Genet. 1975;9:245–284. doi: 10.1146/annurev.ge.09.120175.001333. [DOI] [PubMed] [Google Scholar]
  4. Fox D. P. Some characteristics of the cold hydrolysis technique for staining plant tissues by the feulgen reaction. J Histochem Cytochem. 1969 Apr;17(4):266–272. doi: 10.1177/17.4.266. [DOI] [PubMed] [Google Scholar]
  5. Funderud S., Andreassen R., Haugli F. Size distribution and maturation of newly replicated DNA through the S and G2 phases of Physarum polycephalum. Cell. 1978 Dec;15(4):1519–1526. doi: 10.1016/0092-8674(78)90074-0. [DOI] [PubMed] [Google Scholar]
  6. Funderud S., Haugli F. DNA replication in Physarum polycephalum: characterization of DNA replication products made in vivo in the presence of cycloheximide in strains sensitive and resistant to cycloheximide. Nucleic Acids Res. 1977 Feb;4(2):405–413. doi: 10.1093/nar/4.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. García-Herdugo G., González-Fernández A., López-Sáez J. F. DNA replication in the presence of protein synthesis inhibitors in higher plant cells. Exp Cell Res. 1977 Jan;104(1):1–6. doi: 10.1016/0014-4827(77)90061-1. [DOI] [PubMed] [Google Scholar]
  8. Hand R. Eucaryotic DNA: organization of the genome for replication. Cell. 1978 Oct;15(2):317–325. doi: 10.1016/0092-8674(78)90001-6. [DOI] [PubMed] [Google Scholar]
  9. Hereford L. M., Hartwell L. H. Role of protein synthesis in the replication of yeast DNA. Nat New Biol. 1973 Aug 1;244(135):129–131. doi: 10.1038/newbio244129a0. [DOI] [PubMed] [Google Scholar]
  10. Hori T., Lark K. G. Effect of puromycin on DNA replication in Chinese hamster cells. J Mol Biol. 1973 Jul 5;77(3):391–404. doi: 10.1016/0022-2836(73)90446-4. [DOI] [PubMed] [Google Scholar]
  11. Hyodo M., Koyama H., Ono T. Intermediate fragments of newly replicated DNA in mammalian cells. II. Effect of cycloheximide on DNA chain elongation. Exp Cell Res. 1971 Aug;67(2):461–463. doi: 10.1016/0014-4827(71)90431-9. [DOI] [PubMed] [Google Scholar]
  12. Kowalski J., Cheevers W. P. Synthesis of high molecular weight DNA strands during S phase. J Mol Biol. 1976 Jul 5;104(3):603–615. doi: 10.1016/0022-2836(76)90123-6. [DOI] [PubMed] [Google Scholar]
  13. Lehmann A. R., Ormerod M. G. The replication of DNA in murine lymphoma cells (L5178Y). I. Rate of replication. Biochim Biophys Acta. 1970 Mar 19;204(1):128–143. doi: 10.1016/0005-2787(70)90496-x. [DOI] [PubMed] [Google Scholar]
  14. Lett J. T., Sun C. The production of strand breaks in mammalian DNA by X-rays: at different stages in the cell cycle. Radiat Res. 1970 Dec;44(3):771–787. [PubMed] [Google Scholar]
  15. Muldoon J. J., Evans T. E., Nygaard O. F., Evans H. H. Control of DNA replication by protein synthesis at defined times during the S period in Physarum polycephalum. Biochim Biophys Acta. 1971 Oct 14;247(2):310–321. doi: 10.1016/0005-2787(71)90679-4. [DOI] [PubMed] [Google Scholar]
  16. Schvartzman J. B., Chenet B., Bjerknes C., Van't Hof J. Nascent replicons are synchronously joined at the end of S phase or during G2 phase in peas. Biochim Biophys Acta. 1981 Apr 27;653(2):185–192. doi: 10.1016/0005-2787(81)90154-4. [DOI] [PubMed] [Google Scholar]
  17. Van't Hof J. DNA fiber replication in chromosomes of a higher plant (Pisum sativum). Exp Cell Res. 1975 Jun;93(1):95–104. doi: 10.1016/0014-4827(75)90427-9. [DOI] [PubMed] [Google Scholar]
  18. Walters R. A., Tobey R. A., Hildebrand C. E. Chain elongation and joining of DNA synthesized during hydroxyurea treatment of Chinese hamster cells. Biochim Biophys Acta. 1976 Sep 20;447(1):36–44. doi: 10.1016/0005-2787(76)90093-9. [DOI] [PubMed] [Google Scholar]
  19. Westergaard O., Marcker K. A., Leer J. C. Effect of cycloheximide on maturation of replicative intermediates into high-molecular-weight DNA in Tetrahymena. Eur J Biochem. 1978 May;86(1):255–260. doi: 10.1111/j.1432-1033.1978.tb12306.x. [DOI] [PubMed] [Google Scholar]
  20. Yagura T., Kozu T., Seno T. Arrest of chain growth of replicon-sized intermediates by aphidicolin during rat fibroblast cell chromosome replication. Eur J Biochem. 1982 Mar;123(1):15–21. doi: 10.1111/j.1432-1033.1982.tb06492.x. [DOI] [PubMed] [Google Scholar]

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