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. 1973 Sep 1;58(3):564–573. doi: 10.1083/jcb.58.3.564

INTERMITTENT DNA SYNTHESIS AND PERIODIC EXPRESSION OF ENZYME ACTIVITY IN THE CELL CYCLE OF WI-38

Robert R Klevecz 1, Leon N Kapp 1
PMCID: PMC2109078  PMID: 4795861

Abstract

Synchronous cultures of WI-38 were obtained using an automated system for detachment and partitioning of mitotic cells which operates without the use of inhibitors, altered medium, or lowered temperatures. The generation time in synchronous WI-38 is 19.5 h and the duration of S phase when determined from the percentage of labeled metaphase cells or nuclei is 12 h. DNA replication in WI-38 occurs in three temporally distinct and rapid bursts separated by intervals of greatly reduced synthesis within what is nominally described as the DNA synthetic (S) period. Lactate dehydrogenase (LDH) displayed maxima in G1 between 2 and 4 h and again at 10 and 16 h. Peaks in LDH activity were coordinated with DNA replication in a fashion similar to that reported for diploid Chinese hamster cells. Oscillations in LDH activity are more pronounced in normal diploid fibroblasts than in established and neoplastic lines.

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Selected References

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  1. Balazs L., Schildkraut C. L. DNA replication in synchronized cultured mammalian cells. II. Replication of ribosomal cistrons in thymidine-synchronized HeLa cells. J Mol Biol. 1971 Apr 14;57(1):153–158. doi: 10.1016/0022-2836(71)90125-2. [DOI] [PubMed] [Google Scholar]
  2. Blomquist C. H., Gregg C. T., Tobey R. A. Enzyme and coenzyme levels, oxygen uptake and lactate production in synchronized cultures of Chinese hamster cells. Exp Cell Res. 1971 May;66(1):75–80. doi: 10.1016/s0014-4827(71)80013-7. [DOI] [PubMed] [Google Scholar]
  3. Bosmann H. B., Bernacki R. J. Glycosidase activity. Glycosidase acitivity in L5178Y mouse leukemic cells and the activity of acid phosphatase, beta-galactosidase and beta-N-acetylgalactosaminidase and beta-N-acetylglucosaminidase in a synchronous L5178Y cell population. Exp Cell Res. 1970 Aug;61(2):379–386. doi: 10.1016/0014-4827(70)90461-1. [DOI] [PubMed] [Google Scholar]
  4. Brent T. P. Periodicity of DNA synthetic enzymes during the HeLa cell cycle. Cell Tissue Kinet. 1971 Jul;4(4):297–305. doi: 10.1111/j.1365-2184.1971.tb01541.x. [DOI] [PubMed] [Google Scholar]
  5. CRISTOFALO V. J., KRITCHEVSKY D. GROWTH AND GLYCOLYSIS IN THE HUMAN DIPLOID CELL STRAIN WI-38. Proc Soc Exp Biol Med. 1965 Apr;118:1109–1113. doi: 10.3181/00379727-118-30057. [DOI] [PubMed] [Google Scholar]
  6. Churchill J. R., Studzinski G. P. Thymidine as synchronizing agent. 3. Persistence of cell cycle patterns of phosphatase activities and elevation of nuclease activity during inhibition of DNA synthesis. J Cell Physiol. 1970 Jun;75(3):297–303. doi: 10.1002/jcp.1040750306. [DOI] [PubMed] [Google Scholar]
  7. Comings D. E., Vance C. K. Thermal denaturation of DNA and chromatin of early and late passage human fibroblasts. Gerontologia. 1971;17(2):116–121. doi: 10.1159/000211814. [DOI] [PubMed] [Google Scholar]
  8. Forrest G. L., Klevecz R. R. Synthesis and degradation of microtubule protein in synchronized Chinese hamster cells. J Biol Chem. 1972 May 25;247(10):3147–3152. [PubMed] [Google Scholar]
  9. Gelfant S., Smith J. G., Jr Aging: noncycling cells an explanation. Science. 1972 Oct 27;178(4059):357–361. doi: 10.1126/science.178.4059.357. [DOI] [PubMed] [Google Scholar]
  10. Good P. I. Subcultivations, splits, doublings, and generations in cultures of human diploid fibroblasts. Cell Tissue Kinet. 1972 Jul;5(4):319–323. doi: 10.1111/j.1365-2184.1972.tb00370.x. [DOI] [PubMed] [Google Scholar]
  11. HAYFLICK L., MOORHEAD P. S. The serial cultivation of human diploid cell strains. Exp Cell Res. 1961 Dec;25:585–621. doi: 10.1016/0014-4827(61)90192-6. [DOI] [PubMed] [Google Scholar]
  12. HAYFLICK L. THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS. Exp Cell Res. 1965 Mar;37:614–636. doi: 10.1016/0014-4827(65)90211-9. [DOI] [PubMed] [Google Scholar]
  13. Hinegardner R. T. An improved fluorometric assay for DNA. Anal Biochem. 1971 Jan;39(1):197–201. doi: 10.1016/0003-2697(71)90476-3. [DOI] [PubMed] [Google Scholar]
  14. Howard B. V., Kritchevsky D. The source of cellular lipid in the human diploid cell strain WI-38. Biochim Biophys Acta. 1969 Oct 28;187(3):293–301. doi: 10.1016/0005-2760(69)90002-2. [DOI] [PubMed] [Google Scholar]
  15. Jacobs J. P., Jones C. M., Baille J. P. Characteristics of a human diploid cell designated MRC-5. Nature. 1970 Jul 11;227(5254):168–170. doi: 10.1038/227168a0. [DOI] [PubMed] [Google Scholar]
  16. KISSANE J. M., ROBINS E. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem. 1958 Jul;233(1):184–188. [PubMed] [Google Scholar]
  17. Kanungo M. S., Koul O., Reddy K. R. Concomitant studies on RNA and protein syntheses in tissues of rats of various ages. Exp Gerontol. 1970 Sep;5(3):261–269. doi: 10.1016/0531-5565(70)90047-1. [DOI] [PubMed] [Google Scholar]
  18. Kapp L. N., Okada S. Factors affecting acid phosphatase activity in exponential and synchronized L5178Y mouse leukemia cells. Exp Cell Res. 1972 Jun;72(2):465–472. doi: 10.1016/0014-4827(72)90015-8. [DOI] [PubMed] [Google Scholar]
  19. Klevecz R. R. An automated system for cell cycle analysis. Anal Biochem. 1972 Oct;49(2):407–415. doi: 10.1016/0003-2697(72)90443-5. [DOI] [PubMed] [Google Scholar]
  20. Klevecz R. R., Ruddle F. H. Cyclic changes in enzyme activity in synchronized mammalian cell cultures. Science. 1968 Feb 9;159(3815):634–636. doi: 10.1126/science.159.3815.634. [DOI] [PubMed] [Google Scholar]
  21. Klevecz R. R., Stubblefield E. RNA synthesis in relation to DNA replication in synchronized Chinese hamster cell cultures. J Exp Zool. 1967 Jul;165(2):259–268. doi: 10.1002/jez.1401650210. [DOI] [PubMed] [Google Scholar]
  22. Klevecz R. R. Temporal coordination of DNA replication with enzyme synthesis in diploid and heteroploid cells. Science. 1969 Dec 19;166(3912):1536–1538. doi: 10.1126/science.166.3912.1536. [DOI] [PubMed] [Google Scholar]
  23. Klevecz R. R. Temporal order in mammalian cells. I. The periodic synthesis of lactate dehydrogenase in the cell cycle. J Cell Biol. 1969 Nov;43(2):207–219. doi: 10.1083/jcb.43.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kruse P. F., Jr, Miedema E. Glucose uptake related to proliferation of animal cells in vitro. Proc Soc Exp Biol Med. 1965 Aug-Sep;119(4):1110–1112. doi: 10.3181/00379727-119-30389. [DOI] [PubMed] [Google Scholar]
  25. Lindahl P. E., Sörenby L. A new method for the continuous selection of cells in mitosis. Exp Cell Res. 1966 Sep;43(2):424–434. doi: 10.1016/0014-4827(66)90069-3. [DOI] [PubMed] [Google Scholar]
  26. Macieira-Coelho A., Pontén J., Philipson L. The division cycle and RNA-synthesis in diploid human cells at different passage levels in vitro. Exp Cell Res. 1966 Jun;42(3):673–684. doi: 10.1016/0014-4827(66)90280-1. [DOI] [PubMed] [Google Scholar]
  27. Malamud D. DNA synthesis and the mitotic cycle in frog kidney cells cultivated in vitro. Exp Cell Res. 1967 Feb;45(2):277–280. doi: 10.1016/0014-4827(67)90179-6. [DOI] [PubMed] [Google Scholar]
  28. Melnykovych G., Bishop C. F., Swayze M. A. Fluctuation of alkaline phosphatase activity in synchronized heteroploid cell cultures: effects of prednisolone. J Cell Physiol. 1967 Dec;70(3):231–236. doi: 10.1002/jcp.1040700303. [DOI] [PubMed] [Google Scholar]
  29. Ohara H., Terasima T. Variations of cellular sulfhydryl content during cell cycle of HeLa cells and its correlation to cyclic change of x-ray sensitivity. Exp Cell Res. 1969 Nov;58(1):182–185. doi: 10.1016/0014-4827(69)90133-5. [DOI] [PubMed] [Google Scholar]
  30. Painter R. B., Schaefer A. W. Rate of synthesis along replicons of different kinds of mammalian cells. J Mol Biol. 1969 Nov 14;45(3):467–479. doi: 10.1016/0022-2836(69)90306-4. [DOI] [PubMed] [Google Scholar]
  31. Raff E. C., Houck J. C. Migration and proliferation of diploid human fibroblasts following "wounding" of confluent monolayers. J Cell Physiol. 1969 Dec;74(3):235–244. doi: 10.1002/jcp.1040740304. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Remington J. A., Flickinger R. A. The time of DNA replication in the cell cycle in relation to RNA synthesis in frog embryos. J Cell Physiol. 1971 Jun;77(3):411–422. doi: 10.1002/jcp.1040770316. [DOI] [PubMed] [Google Scholar]
  34. Remington J. A., Klevecz R. R. Families of replicating units in cultured hamster fibroblasts. Exp Cell Res. 1973 Feb;76(2):410–418. doi: 10.1016/0014-4827(73)90393-5. [DOI] [PubMed] [Google Scholar]
  35. Ryan J. M., Cristofalo V. J. Histone acetylation during aging of human cells in culture. Biochem Biophys Res Commun. 1972 Aug 21;48(4):735–742. doi: 10.1016/0006-291x(72)90668-7. [DOI] [PubMed] [Google Scholar]
  36. SOUKUPOVA M., HOLECKOVA E. THE LATENT PERIOD OF EXPLANTED ORGANS OF NEWBORN, ADULT AND SENILE RATS. Exp Cell Res. 1964 Feb;33:361–367. doi: 10.1016/0014-4827(64)90001-1. [DOI] [PubMed] [Google Scholar]
  37. Stambrook P. J., Flickinger R. A. Changes i chromosomal DNA replication tterns in developing frog embryos. J Exp Zool. 1970 May;174(1):101–113. doi: 10.1002/jez.1401740110. [DOI] [PubMed] [Google Scholar]
  38. TODARO G. J., GREEN H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol. 1963 May;17:299–313. doi: 10.1083/jcb.17.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tobey R. A., Crissman H. A., Kraemer P. M. A method for comparing effects of different synchronizing protocols on mammalian cell cycle traverse. The traverse perturbation index. J Cell Biol. 1972 Sep;54(3):638–645. doi: 10.1083/jcb.54.3.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wiebel F., Baserga R. Early alterations in amino acid pools and protein synthesis of diploid fibroblasts stimulated to synthesize DNA by addition of serum. J Cell Physiol. 1969 Oct;74(2):191–202. doi: 10.1002/jcp.1040740211. [DOI] [PubMed] [Google Scholar]
  41. Willingham M. C., Johnson G. S., Pastan I. Control of DNA synthesis and mitosis in 3T3 cells by cyclic AMP. Biochem Biophys Res Commun. 1972 Aug 21;48(4):743–748. doi: 10.1016/0006-291x(72)90669-9. [DOI] [PubMed] [Google Scholar]

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