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Identification of Proteins Whose Synthesis Is Modulated During the Cell Cycle of Saccharomyces cerevisiae

机译：鉴定啤酒酵母细胞周期中其合成受到调控的蛋白质

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We examined the synthesis and turnover of individual proteins in the Saccharomyces cerevisiae cell cycle. Proteins were pulse-labeled with radioactive isotope (35S or 14C) in cells at discrete cycle stages and then resolved on two-dimensional gels and analyzed by a semiautomatic procedure for quantitating gel electropherogram-autoradiographs. The cells were obtained by one of three methods: (i) isolation of synchronous subpopulations of growing cells by zonal centrifugation; (ii) fractionation of pulse-labeled steady-state cultures according to cell age; and (iii) synchronization of cells with the mating pheromone, α-factor. In confirmation of previous studies, we found that the histones H4, H2A, and H2B were synthesized almost exclusively in the late G1 and early S phases. In addition, we identified eight proteins whose rates of synthesis were modulated in the cell cycle, and nine proteins (of which five, which may well be related, were unstable, with half-lives of 10 to 15 min) that might be regulated in the cell cycle by periodic synthesis, modification, or degradation. Based on the time of maximal labeling in the cell cycle and on experiments with α-factor and hydroxyurea, we assigned the cell cycle proteins to two classes: proteins in class I were labeled principally in early G1 phase and at a late stage of the cycle, whereas those in class II were primarily synthesized at times ranging from late G1 to mid S phase. At least one major control point for the cell cycle proteins occurred between “start” and early S phase. A set of stress-responsive proteins was also identified and analyzed. The rates of synthesis of these proteins were affected by certain perturbations that resulted during selection of synchronous cell populations and by heat shock.
机译：我们研究了酿酒酵母（Saccharomyces cerevisiae）细胞周期中单个蛋白质的合成和更新。在细胞的离散周期中用放射性同位素（ 35 S或 14 C）对蛋白质进行脉冲标记，然后在二维凝胶上分离，并通过半自动程序进行分析。定量凝胶电泳图-放射自显影仪。通过以下三种方法之一获得细胞：（i）通过区域离心分离生长细胞的同步亚群; （ii）根据细胞年龄分级分离脉冲标记的稳态培养物; （iii）细胞与交配信息素α因子同步。为了证实先前的研究，我们发现组蛋白H4，H2A和H2B几乎完全在G1晚期和S早期阶段合成。此外，我们鉴定了8​​种蛋白质，这些蛋白质的合成速率在细胞周期中受到调节，并且9种蛋白质（其中5种可能很相关，它们是不稳定的，半衰期为10至15分钟）可能在细胞周期中受到调节。通过周期性的合成，修饰或降解来改变细胞周期。根据细胞周期中最大标记的时间以及α-因子和羟基脲的实验，我们将细胞周期蛋白分为两类：I类蛋白主要在G1早期和周期后期进行标记。 ，而II类中的那些主要是从G1晚期到S中期的时间合成的。细胞周期蛋白的至少一个主要控制点发生在“开始”和早期S期之间。还鉴定并分析了一组应激反应蛋白。这些蛋白质的合成速率受同步细胞群选择过程中产生的某些扰动以及热激影响。

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