Uptake of amino acids and thymidine during the first cell cycle of synchronized hamster cells

The net total uptake of four amino acids (valine, leucine, lysine and methionine) used at concentrations required for growth, and of thymidine at tracer concentrations, has been studied during the first cell cycle of an asparagine-dependent strain of transformed BHK cells synchronized by asparagine starvation. The rate of the total uptake of the amino acids, the free pool of the amino acids taken up, and the rate of their incorporation into protein at the end of the first cell cycle were, on the average, 12-fold that at the beginning of the cell cycle. The increase in these parameters during the cell cycle was not linear. The uptake of thymidine started before the onset of DNA synthesis and proceeded linearly beyond the peak of the S phase. The rate of accumulation of thymidine into the acid-soluble fraction also increased during the S phase, apart from a tendency to plateau off at the peak of this phase. It reached a second plateau towards the end of the cell cycle, and then declined slightly. Evidence is presented which suggests that the total quantity of protein synthesized during the cell cycle is more than the newly synthesized protein present in the cells at the end of the cell cycle; this indicated degradation and/or secretion of a substantial proportion of the newly synthesized protein. The total protein synthesized at different time points in the cell cycle appeared to contain different proportions of the amino acids used.