Elliott S G, McLaughlin C S
J Bacteriol. 1979 Mar;137(3):1185-90. doi: 10.1128/jb.137.3.1185-1190.1979.
We have used a novel technique to study the synthesis, modification and degradation of proteins during the cell cycle in Saccharomyces cerevisiae. Logarithmically growing cells were pulse-labeled twice, with the pulses separated in time by more than one generation. Subsequently, the cells were fractionated as to their position in the cell cycle by centrifugal elutriation, and for different proteins the ratio of radioactive material from the two pulses was then determined. Periodic degradation, synthesis, or modification would produce periodic variations in the ratio of counts. Two-dimensional gel electrophoresis was used to examine 110 different proteins at different times of the cell cycle. All but two proteins had a constant ratio of counts through the cell cycle. This indicates that the rate of synthesis of individual proteins increases exponentially during the cell cycle and that periodic degradation or modification of proteins is not a general feature of the cell cycle in S. cerevisiae.
我们采用了一种新技术来研究酿酒酵母细胞周期中蛋白质的合成、修饰和降解过程。对数生长期的细胞进行了两次脉冲标记,两次脉冲之间的时间间隔超过一个细胞世代。随后,通过离心淘析根据细胞在细胞周期中的位置对细胞进行分级分离,然后测定不同蛋白质来自两次脉冲的放射性物质的比例。周期性的降解、合成或修饰会导致计数比例出现周期性变化。利用二维凝胶电泳在细胞周期的不同时间检测了110种不同的蛋白质。除两种蛋白质外,所有蛋白质在整个细胞周期中的计数比例都保持恒定。这表明在细胞周期中单个蛋白质的合成速率呈指数增长,并且蛋白质的周期性降解或修饰并非酿酒酵母细胞周期的普遍特征。