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. 1987 Dec;84(24):9025–9029. doi: 10.1073/pnas.84.24.9025

Periodic mitotic events induced in the absence of DNA replication.

R Schlegel 1, A B Pardee 1
PMCID: PMC299684  PMID: 3480528

Abstract

We have discovered and report here a means of separating a mitotic "subcycle" from the G1- and S-phase events of the mammalian cell cycle. Time-lapse videomicroscopy of Syrian hamster fibroblast (BHK) cells revealed that caffeine could induce multiple entries into mitosis while cells were blocked in DNA synthesis. As with normal mitoses, the abundance of mitosis-specific phosphoproteins was coupled with the condensation of chromatin. The BHK temperature-sensitive mutant tsBN2 also completed multiple entries into mitosis while arrested during DNA replication and raised to the restrictive temperature. Periodic mitotic events occurred even when BHK cells were exposed to low concentrations of serum or cycloheximide, conditions that prevent the cycling of BHK cells by blocking their entry into S phase. These results suggest that an oscillation governing the activation and inactivation of mitotic factors can be generated in mammalian cells and uncoupled from the G1 and DNA replication events of the normal cell cycle. This system will be useful for examining the molecular nature of mitotic factors.

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Selected References

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

  1. Adlakha R. C., Sahasrabuddhe C. G., Wright D. A., Rao P. N. Evidence for the presence of inhibitors of mitotic factors during G1 period in mammalian cells. J Cell Biol. 1983 Dec;97(6):1707–1713. doi: 10.1083/jcb.97.6.1707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carter S. B. Effects of cytochalasins on mammalian cells. Nature. 1967 Jan 21;213(5073):261–264. doi: 10.1038/213261a0. [DOI] [PubMed] [Google Scholar]
  3. Davis F. M., Tsao T. Y., Fowler S. K., Rao P. N. Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2926–2930. doi: 10.1073/pnas.80.10.2926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Evans T., Rosenthal E. T., Youngblom J., Distel D., Hunt T. Cyclin: a protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division. Cell. 1983 Jun;33(2):389–396. doi: 10.1016/0092-8674(83)90420-8. [DOI] [PubMed] [Google Scholar]
  5. GROSS P. R., MALKIN L. I., MOYER W. A. TEMPLATES FOR THE FIRST PROTEINS OF EMBRYONIC DEVELOPMENT. Proc Natl Acad Sci U S A. 1964 Mar;51:407–414. doi: 10.1073/pnas.51.3.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gerhart J., Wu M., Kirschner M. Cell cycle dynamics of an M-phase-specific cytoplasmic factor in Xenopus laevis oocytes and eggs. J Cell Biol. 1984 Apr;98(4):1247–1255. doi: 10.1083/jcb.98.4.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Halleck M. S., Lumley-Sapanski K., Reed J. A., Iyer A. P., Mastro A. M., Schlegel R. A. Characterization of protein kinases in mitotic and meiotic cell extracts. FEBS Lett. 1984 Feb 27;167(2):193–198. doi: 10.1016/0014-5793(84)80125-8. [DOI] [PubMed] [Google Scholar]
  8. Hamlin J. L., Pardee A. B. S phase synchrony in monolayer CHO cultures. Exp Cell Res. 1976 Jul;100(2):265–275. doi: 10.1016/0014-4827(76)90147-6. [DOI] [PubMed] [Google Scholar]
  9. KISHIMOTO S., LIEBERMAN I. SYNTHESIS OF RNA AND PROTEIN REQUIRED FOR THE MITOSIS OF MAMMALIAN CELLS. Exp Cell Res. 1964 Oct;36:92–101. doi: 10.1016/0014-4827(64)90163-6. [DOI] [PubMed] [Google Scholar]
  10. Krishan A. Cytochalasin-B: time-lapse cinematographic studies on its effects on cytokinesis. J Cell Biol. 1972 Sep;54(3):657–664. doi: 10.1083/jcb.54.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Newport J. W., Kirschner M. W. Regulation of the cell cycle during early Xenopus development. Cell. 1984 Jul;37(3):731–742. doi: 10.1016/0092-8674(84)90409-4. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Nishimoto T., Ishida R., Ajiro K., Yamamoto S., Takahashi T. The synthesis of protein(s) for chromosome condensation may be regulated by a post-transcriptional mechanism. J Cell Physiol. 1981 Nov;109(2):299–308. doi: 10.1002/jcp.1041090213. [DOI] [PubMed] [Google Scholar]
  14. Osmani S. A., May G. S., Morris N. R. Regulation of the mRNA levels of nimA, a gene required for the G2-M transition in Aspergillus nidulans. J Cell Biol. 1987 Jun;104(6):1495–1504. doi: 10.1083/jcb.104.6.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Rosenthal E. T., Hunt T., Ruderman J. V. Selective translation of mRNA controls the pattern of protein synthesis during early development of the surf clam, Spisula solidissima. Cell. 1980 Jun;20(2):487–494. doi: 10.1016/0092-8674(80)90635-2. [DOI] [PubMed] [Google Scholar]
  17. Russell P., Nurse P. Negative regulation of mitosis by wee1+, a gene encoding a protein kinase homolog. Cell. 1987 May 22;49(4):559–567. doi: 10.1016/0092-8674(87)90458-2. [DOI] [PubMed] [Google Scholar]
  18. Russell P., Nurse P. The mitotic inducer nim1+ functions in a regulatory network of protein kinase homologs controlling the initiation of mitosis. Cell. 1987 May 22;49(4):569–576. doi: 10.1016/0092-8674(87)90459-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Simanis V., Nurse P. The cell cycle control gene cdc2+ of fission yeast encodes a protein kinase potentially regulated by phosphorylation. Cell. 1986 Apr 25;45(2):261–268. doi: 10.1016/0092-8674(86)90390-9. [DOI] [PubMed] [Google Scholar]
  22. Smith L. D., Ecker R. E. Protein synthesis in enucleated eggs of Rana pipiens. Science. 1965 Nov 5;150(3697):777–779. doi: 10.1126/science.150.3697.777. [DOI] [PubMed] [Google Scholar]
  23. Sunkara P. S., Wright D. A., Rao P. N. Mitotic factors from mammalian cells induce germinal vesicle breakdown and chromosome condensation in amphibian oocytes. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2799–2802. doi: 10.1073/pnas.76.6.2799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Swenson K. I., Farrell K. M., Ruderman J. V. The clam embryo protein cyclin A induces entry into M phase and the resumption of meiosis in Xenopus oocytes. Cell. 1986 Dec 26;47(6):861–870. doi: 10.1016/0092-8674(86)90801-9. [DOI] [PubMed] [Google Scholar]
  25. Tobey R. A., Crissman H. A. Preparation of large quantities of synchronized mammalian cells in late G1 in the pre-DNA replicative phase of the cell cycle. Exp Cell Res. 1972 Dec;75(2):460–464. doi: 10.1016/0014-4827(72)90453-3. [DOI] [PubMed] [Google Scholar]
  26. Tobey R. A., Petersen D. F., Anderson E. C., Puck T. T. Life cycle analysis of mammalian cells. 3. The inhibition of division in Chinese hamster cells by puromycin and actinomycin. Biophys J. 1966 Sep;6(5):567–581. doi: 10.1016/s0006-3495(66)86678-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wasserman W. J., Masui Y. Effects of cyclohexamide on a cytoplasmic factor initiating meiotic naturation in Xenopus oocytes. Exp Cell Res. 1975 Mar 15;91(2):381–388. doi: 10.1016/0014-4827(75)90118-4. [DOI] [PubMed] [Google Scholar]
  28. Woodford T. A., Pardee A. B. Histone H1 kinase in exponential and synchronous populations of Chinese hamster fibroblasts. J Biol Chem. 1986 Apr 5;261(10):4669–4676. [PubMed] [Google Scholar]

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