Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1990 Jun 1;110(6):1855–1859. doi: 10.1083/jcb.110.6.1855

Caffeine overcomes a restriction point associated with DNA replication, but does not accelerate mitosis

PMCID: PMC2116112  PMID: 2161852

Abstract

Mitotic chromosome condensation is normally dependent on the previous completion of replication. Caffeine spectacularly deranges cell cycle controls after DNA polymerase inhibition or DNA damage; it induces the condensation, in cells that have not completed replication, of fragmented nuclear structures, analogous to the S-phase prematurely condensed chromosomes seen when replicating cells are fused with mitotic cells. Caffeine has been reported to induce S-phase condensation in cells where replication is arrested, by accelerating cell cycle progression as well as by uncoupling it from replication; for, in BHK or CHO hamster cells arrested in early S-phase and given caffeine, condensed chromosomes appear well before the normal time at which mitosis occurs in cells released from arrest. However, we have found that this apparent acceleration depends on the technique of synchrony and cell line employed. In other cells, and in synchronized hamster cells where the cycle has not been subjected to prolonged continual arrest, condensation in replication-arrested cells given caffeine occurs at the same time as normal mitosis in parallel populations where replication is allowed to proceed. This caffeine- induced condensation is therefore "premature" with respect to the chromatin structure of the S-phase nucleus, but not with respect to the timing of the normal cycle. Caffeine in replication-arrested cells thus overcomes the restriction on the formation of mitotic condensing factors that is normally imposed during DNA replication, but does not accelerate the timing of condensation unless cycle controls have previously been disturbed by synchronization procedures.

Full Text

The Full Text of this article is available as a PDF (494.6 KB).

Selected References

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

  1. Brinkley B. R., Zinkowski R. P., Mollon W. L., Davis F. M., Pisegna M. A., Pershouse M., Rao P. N. Movement and segregation of kinetochores experimentally detached from mammalian chromosomes. Nature. 1988 Nov 17;336(6196):251–254. doi: 10.1038/336251a0. [DOI] [PubMed] [Google Scholar]
  2. Busse P. M., Bose S. K., Jones R. W., Tolmach L. J. The action of caffeine on X-irradiated HeLa cells. III. Enhancement of X-ray-induced killing during G2 arrest. Radiat Res. 1978 Nov;76(2):292–307. [PubMed] [Google Scholar]
  3. 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]
  4. Creasey D. C., Ts'o P. O. DNA replication in Syrian hamster cells transiently exposed to hydroxyurea. Cancer Res. 1988 Nov 15;48(22):6298–6302. [PubMed] [Google Scholar]
  5. Dolbeare F., Gratzner H., Pallavicini M. G., Gray J. W. Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5573–5577. doi: 10.1073/pnas.80.18.5573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fallon R. J., Cox R. P. Cell cycle analysis of sodium butyrate and hydroxyurea, inducers of ectopic hormone production in HeLa cells. J Cell Physiol. 1979 Aug;100(2):251–262. doi: 10.1002/jcp.1041000206. [DOI] [PubMed] [Google Scholar]
  7. Hahn P., Kapp L. N., Morgan W. F., Painter R. B. Chromosomal changes without DNA overproduction in hydroxyurea-treated mammalian cells: implications for gene amplification. Cancer Res. 1986 Sep;46(9):4607–4612. [PubMed] [Google Scholar]
  8. Johnson R. T., Rao P. N. Mammalian cell fusion: induction of premature chromosome condensation in interphase nuclei. Nature. 1970 May 23;226(5247):717–722. doi: 10.1038/226717a0. [DOI] [PubMed] [Google Scholar]
  9. Kimelman D., Kirschner M., Scherson T. The events of the midblastula transition in Xenopus are regulated by changes in the cell cycle. Cell. 1987 Feb 13;48(3):399–407. doi: 10.1016/0092-8674(87)90191-7. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Lau C. C., Pardee A. B. Mechanism by which caffeine potentiates lethality of nitrogen mustard. Proc Natl Acad Sci U S A. 1982 May;79(9):2942–2946. doi: 10.1073/pnas.79.9.2942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mariani B. D., Schimke R. T. Gene amplification in a single cell cycle in Chinese hamster ovary cells. J Biol Chem. 1984 Feb 10;259(3):1901–1910. [PubMed] [Google Scholar]
  13. Mullinger A. M., Johnson R. T. Units of chromosome replication and packing. J Cell Sci. 1983 Nov;64:179–193. doi: 10.1242/jcs.64.1.179. [DOI] [PubMed] [Google Scholar]
  14. Murray A. W. Cell biology: the cell cycle as a cdc2 cycle. Nature. 1989 Nov 2;342(6245):14–15. doi: 10.1038/342014a0. [DOI] [PubMed] [Google Scholar]
  15. Musk S. R., Downes C. S., Johnson R. T. Caffeine induces uncoordinated expression of cell cycle functions after ultraviolet irradiation. Accelerated cycle transit, sister chromatid exchanges and premature chromosome condensation in a transformed Indian muntjac cell line. J Cell Sci. 1988 Aug;90(Pt 4):591–599. doi: 10.1242/jcs.90.4.591. [DOI] [PubMed] [Google Scholar]
  16. Nishimoto T., Eilen E., Basilico C. Premature of chromosome condensation in a ts DNA- mutant of BHK cells. Cell. 1978 Oct;15(2):475–483. doi: 10.1016/0092-8674(78)90017-x. [DOI] [PubMed] [Google Scholar]
  17. Ohtsubo M., Kai R., Furuno N., Sekiguchi T., Sekiguchi M., Hayashida H., Kuma K., Miyata T., Fukushige S., Murotsu T. Isolation and characterization of the active cDNA of the human cell cycle gene (RCC1) involved in the regulation of onset of chromosome condensation. Genes Dev. 1987 Aug;1(6):585–593. doi: 10.1101/gad.1.6.585. [DOI] [PubMed] [Google Scholar]
  18. Painter R. B. Effect of caffeine on DNA synthesis in irradiated and unirradiated mammalian cells. J Mol Biol. 1980 Nov 5;143(3):289–301. doi: 10.1016/0022-2836(80)90191-6. [DOI] [PubMed] [Google Scholar]
  19. Pillidge L., Downes C. S., Johnson R. T. Defective post-replication recovery and u.v. sensitivity in a simian virus 40-transformed Indian muntjac cell line. Int J Radiat Biol Relat Stud Phys Chem Med. 1986 Jul;50(1):119–136. doi: 10.1080/09553008614550501. [DOI] [PubMed] [Google Scholar]
  20. Raff J. W., Glover D. M. Nuclear and cytoplasmic mitotic cycles continue in Drosophila embryos in which DNA synthesis is inhibited with aphidicolin. J Cell Biol. 1988 Dec;107(6 Pt 1):2009–2019. doi: 10.1083/jcb.107.6.2009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rowley R. Is G2-arrest an active cellular response to irradiation? Int J Radiat Biol Relat Stud Phys Chem Med. 1985 Nov;48(5):811–820. doi: 10.1080/09553008514551911. [DOI] [PubMed] [Google Scholar]
  22. Sasaki H., Nishimoto T. Chromosome condensation may enhance X-ray-related cell lethality in a temperature-sensitive mutant (tsBN2) of baby hamster kidney cells (BHK21). Radiat Res. 1987 Mar;109(3):407–418. [PubMed] [Google Scholar]
  23. Schlegel R., Croy R. G., Pardee A. B. Exposure to caffeine and suppression of DNA replication combine to stabilize the proteins and RNA required for premature mitotic events. J Cell Physiol. 1987 Apr;131(1):85–91. doi: 10.1002/jcp.1041310113. [DOI] [PubMed] [Google Scholar]
  24. Schlegel R., Pardee A. B. Caffeine-induced uncoupling of mitosis from the completion of DNA replication in mammalian cells. Science. 1986 Jun 6;232(4755):1264–1266. doi: 10.1126/science.2422760. [DOI] [PubMed] [Google Scholar]
  25. Schlegel R., Pardee A. B. Periodic mitotic events induced in the absence of DNA replication. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9025–9029. doi: 10.1073/pnas.84.24.9025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Walters R. A., Gurley L. R., Tobey R. A. Effects of caffeine on radiation-induced phenomena associated with cell-cycle traverse of mammalian cells. Biophys J. 1974 Feb;14(2):99–118. doi: 10.1016/S0006-3495(74)70002-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Watson J. V. Dual laser beam focussing for flow cytometry through a single crossed cylindrical lens pair. Cytometry. 1981 Jul;2(1):14–19. doi: 10.1002/cyto.990020103. [DOI] [PubMed] [Google Scholar]
  28. Watson J. V., Horsnell T. S., Smith P. J. Data compression: 8-dimensional flow cytometric data processing with 28K addressable computer memory. J Immunol Methods. 1988 Oct 26;113(2):205–214. doi: 10.1016/0022-1759(88)90333-x. [DOI] [PubMed] [Google Scholar]
  29. Watson J. V., Sikora K., Evan G. I. A simultaneous flow cytometric assay for c-myc oncoprotein and DNA in nuclei from paraffin embedded material. J Immunol Methods. 1985 Oct 24;83(1):179–192. doi: 10.1016/0022-1759(85)90071-7. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES