Adaptation of Thymidine Utilization to Changing Rates of DNA Synthesis in the Cell Cycle

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 1979 Sep 28

PMID 117296

Citations 1

Authors

G G Miller

J C Schaer

J R Gautschi

R Schindler

Affiliations

Soon will be listed here.

Abstract

In synchronous cultures of P-815 murine mastocytoma and of Chinese hamster ovary (CHO) cells, the relative contribution of exogenous thymidine to DNA synthesis was studied by comparing rates of (3H)thymidine incorporation with the rate of DNA synthesis as derived from incorporation of (3H)thymidine (10(-5) M) in the presence of amethopterin. In synchronous P-815 cultures, time-dependent variations of DNA synthesis rates were in close agreement with those of (3H)thymidine incorporation rates at concentrations of the precursor ranging from 5 x 10(-8) to 10(-5) M. Similarly, in synchronous CHO cell cultures prepared by two different methods, time-dependent changes in DNA synthesis rate were almost identical with those of the rate of incorporation of (3H)thymidine supplied at 5 x 10(-8) M. Thus, at a given thymidine concentration in the medium, the proportion of thymine residues in DNA that were derived from exogenous thymidine remained nearly constant, even though rates of cellular DNA synthesis underwent pronounced changes. This indicates that in the synchronous culture systems used, utilization of exogenous thymidine is efficiently adapted to changing rates of DNA synthesis.

Citing Articles

Effect of arabinosyl cytosine on the level of DNA polymerase and thymidine kinase activity in PHA-stimulated human tonsillar lymphocytes.

Staub M, Sasvari-Szekely M, Spasokukockaja T, Antoni F Mol Cell Biochem. 1982; 42(1):37-44.

PMID: 7062908 DOI: 10.1007/BF00223537.

References

Tobey R, Anderson E, Petersen D . Properties of mitotic cells prepared by mechanically shaking monolayer cultures of Chinese hamster cells. J Cell Physiol. 1967; 70(1):63-8. DOI: 10.1002/jcp.1040700109. View

Tobia A, SCHILDKRAUT C, MAIO J . Deoxyribonucleic acid replication in synchronized cultured mammalian cells. I. Time of synthesis of molecules of different average uanine + cytosine content. J Mol Biol. 1970; 54(3):499-515. DOI: 10.1016/0022-2836(70)90122-1. View

Housman D, Huberman J . Changes in the rate of DNA replication fork movement during S phase in mammalian cells. J Mol Biol. 1975; 94(2):173-81. DOI: 10.1016/0022-2836(75)90076-5. View

Plagemann P, Richey D, Zylka J, Erbe J . Thymidine transport by Novikoff rat hepatoma cells synchronized by double hydroxyurea treatment. Exp Cell Res. 1974; 83(2):303-10. DOI: 10.1016/0014-4827(74)90343-7. View

SCHILDKRAUT C, MARMUR J, Doty P . Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. J Mol Biol. 1962; 4:430-43. DOI: 10.1016/s0022-2836(62)80100-4. View

Dendy P, Cleaver J . AN INVESTIGATION OF (A) VARIATION IN RATE OF DNA-SYNTHESIS DURING S-PHASE IN MOUSE L-CELLS; (B) EFFECT OF ULTRA-VIOLET RADIATION ON RATE OF DNA-SYNTHESIS. Int J Radiat Biol Relat Stud Phys Chem Med. 1964; 8:301-15. DOI: 10.1080/09553006414550371. View

Sinclair W . Hydroxyurea: differential lethal effects on cultured mammalian cells during the cell cycle. Science. 1965; 150(3704):1729-31. DOI: 10.1126/science.150.3704.1729. View

Meyn R, Hewitt R, HUMPHREY R . Evaluation of S phase synchronization by analysis of DNA replication in 5-bromodeoxyuridine. Exp Cell Res. 1973; 82(1):137-42. DOI: 10.1016/0014-4827(73)90255-3. View

Siegers M, Schaer J, Hirsiger H, Schindler R . Determination of rates of DNA synthesis in cultured mammalian cell populations. J Cell Biol. 1974; 62(2):305-15. PMC: 2109404. DOI: 10.1083/jcb.62.2.305. View

10.

Bostock C, Prescott D . Buoyant density of DNA synthesized at different stages of S phase in Chinese hamster cells. Exp Cell Res. 1971; 64(2):481-4. DOI: 10.1016/0014-4827(71)90106-6. View

11.

TERASIMA T, Tolmach L . Growth and nucleic acid synthesis in synchronously dividing populations of HeLa cells. Exp Cell Res. 1963; 30:344-62. DOI: 10.1016/0014-4827(63)90306-9. View

12.

Painter R, Schaefer A . Variation in the rate of DNA chain growth through the S phase in HeLa cells. J Mol Biol. 1971; 58(1):289-95. DOI: 10.1016/0022-2836(71)90247-6. View

13.

Schaer J, Schindler R . The requirement of mammalian cell cultures for serum proteins. Growth-promoting activity of pepsin-digested serum albumin in different media. Biochim Biophys Acta. 1967; 147(1):154-61. DOI: 10.1016/0005-2795(67)90098-0. View

14.

Schaer J, Ramseier L, Schindler R . Studies on the division cycle of mammalian cells. IV. Incorporation of labeled precursors into DNA of synchronously dividing cells in culture. Exp Cell Res. 1971; 65(1):17-22. DOI: 10.1016/s0014-4827(71)80044-7. View

15.

Klevecz R, Keniston B . The temporal structure of S phase. Cell. 1975; 5(2):195-203. DOI: 10.1016/0092-8674(75)90027-6. View

16.

Sordat M, Sordat B, COTTIER H, Hess M, Riedwyl H, Chanana A . Studies on lymphocytes. XV. Analysis of the in vivo division cycle of large lymphoid cells in calf thoracic duct using combined microspectrophotometry and autoradiography. Exp Cell Res. 1972; 70(1):145-53. DOI: 10.1016/0014-4827(72)90192-9. View

17.

Schaer J, Maurer U, Schindler R . Determination of thymidine in serum used for cell culture media. Exp Cell Biol. 1978; 46(1-2):1-10. DOI: 10.1159/000162879. View

18.

Sueoka N, MARMUR J, DOTY 2nd P . Dependence of the density of deoxyribonucleic acids on guanine-cytosine content. Nature. 1959; 183(4673):1429-31. DOI: 10.1038/1831429a0. View

19.

PUCK T, CIECIURA S, Fisher H . Clonal growth in vitro of human cells with fibroblastic morphology; comparison of growth and genetic characteristics of single epithelioid and fibroblast-like cells from a variety of human organs. J Exp Med. 1957; 106(1):145-58. PMC: 2136737. DOI: 10.1084/jem.106.1.145. View

20.

Hanania N, Caneva R, Tapiero H, Harel J . Distribution of repetitious DNA in randomly growing and synchronized Chinese hamster cells. Exp Cell Res. 1975; 90(1):79-86. DOI: 10.1016/0014-4827(75)90359-6. View