DETERMINATION OF RATES OF DNA SYNTHESIS IN CULTURED MAMMALIAN CELL POPULATIONS
Open Access
- 1 August 1974
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 62 (2), 305-315
- https://doi.org/10.1083/jcb.62.2.305
Abstract
In cultures of a murine mastocytoma, endogenous synthesis of thymidine phosphates, as determined by the incorporation of [(3)H]deoxyuridine into DNA, was reduced within 15 min to less than 3% of control values by the addition of amethopterin (10 microM) in combination with hypoxanthine and glycine. If [(3)H]thymidine and unlabeled thymidine were added simultaneously with amethopterin, the increase with time of radioactivity in cellular DNA was linear at least between 30 and 90 min, while radioactivity in the acid-soluble nucleotide fraction remained constant during this time interval, indicating that intracellular thymidine nucleotides had the same specific activity as exogenously supplied [(3)H]thymidine. This permitted calculation of the amount of thymidine incorporated per hour into DNA of 10(6) cells. In conjunction with the base composition of mouse DNA, these results were used to calculate rates of DNA synthesis. Cell proliferation rate, cell cycle time, and the duration of the S period were not affected to any appreciable extent by the addition of amethopterin and thymidine. Rates of DNA synthesis, as derived from thymidine incorporation rates, were in good agreement with those derived from the measured mean DNA content of exponentially multiplying cells and rates of cell proliferation.Keywords
This publication has 18 references indexed in Scilit:
- THYMIDINE TRANSPORT BY CULTURED NOVIKOFF HEPATOMA CELLS AND UPTAKE BY SIMPLE DIFFUSION AND RELATIONSHIP TO INCORPORATION INTO DEOXYRIBONUCLEIC ACIDThe Journal of cell biology, 1972
- STUDIES ON THE DIVISION CYCLE OF MAMMALIAN CELLSThe Journal of cell biology, 1971
- Studies on the division cycle of mammalian cells: IV. Incorporation of labeled precursors into DNA of synchronously dividing cells in cultureExperimental Cell Research, 1971
- Discrepancies between precursor uptake and DNA synthesis in mammalian cellsJournal of Cellular Physiology, 1969
- The effect of endogenous pools of thymidylate on the apparent rate of DNA synthesisExperimental Cell Research, 1969
- Causal relationship between completion of DNA synthesis and onset of the G2 period in the division cycleEuropean Journal of Cancer (1965), 1967
- Studies concerning inhibition of the synthesis of deoxycytidine by phosphorylated derivatives of thymidineBiochimica et Biophysica Acta, 1960
- Cell population kinetics in the intestinal epithelium of the mouseExperimental Cell Research, 1959
- Conservation of desoxyribonucleic acid phosphorus in animal cells propagated in vitroBiochimica et Biophysica Acta, 1956
- Metabolic stability of DNA in fibroblast culturesBiochimica et Biophysica Acta, 1956