Creanor J, Mitchison J M
J Cell Sci. 1982 Dec;58:263-85. doi: 10.1242/jcs.58.1.263.
The rate of protein synthesis through the cell cycle of Schizosaccharomyces pombe has been determined from the incorporation of pulses of [3H]tryptophan in synchronous cultures prepared by selection in an elutriating rotor. This selection procedure caused minimal perturbations as judged by asynchronous control cultures, which had also been put through the rotor. The rate of synthesis showed a periodic pattern rather than a smooth exponential increase. There was a sharp increase in the rate at an 'acceleration point' at about 0.9 of the cycle. Model-fitting by a novel procedure suggests that the average single cell has an increasing rate of protein synthesis for the first 60% of the cycle and a constant rate for the remaining 40%. The same pattern was shown in less extensive experiments with [3H]leucine and [3H]phenylalanine. It was also shown in a series of size mutants, which indicates that the pattern is not size-related, in contrast to earlier work on the rates of synthesis of messenger RNA. However, one large mutant (cdc 2.M35r20) had a significantly earlier acceleration point. Care was taken to justify the assumption that the rate of incorporation of tryptophan was a valid measure of the rate of protein synthesis. A tryptophan auxotroph was used to eliminate the problem of endogenous supply and the size of the metabolic pool was measured through the cycle. This pool did not show cell-cycle related fluctuations. An operational model of the pools is presented.
通过在淘洗转子中进行选择制备同步培养物,利用[3H]色氨酸脉冲掺入法测定了粟酒裂殖酵母细胞周期中的蛋白质合成速率。通过异步对照培养物判断,这种选择程序造成的扰动最小,异步对照培养物也经过了转子处理。合成速率呈现出周期性模式,而非平稳的指数增长。在周期约0.9处的“加速点”,合成速率急剧增加。一种新方法的模型拟合表明,平均单细胞在周期的前60%蛋白质合成速率不断增加,在剩余40%的周期内速率恒定。用[3H]亮氨酸和[3H]苯丙氨酸进行的规模较小的实验也显示出相同模式。在一系列大小突变体中也观察到了这种模式,这表明该模式与细胞大小无关,这与早期关于信使核糖核酸合成速率的研究结果相反。然而,一个大的突变体(cdc 2.M35r20)的加速点明显更早。已谨慎证明色氨酸掺入速率是蛋白质合成速率有效指标这一假设的合理性。使用色氨酸营养缺陷型来消除内源性供应问题,并在整个周期中测量代谢池的大小。该代谢池未显示出与细胞周期相关的波动。本文提出了代谢池的操作模型。