Godon C, Lagniel G, Lee J, Buhler J M, Kieffer S, Perrot M, Boucherie H, Toledano M B, Labarre J
Service de Biochimie et Génétique Moléculaire, CEA-Saclay, F-91191 Gif-sur-Yvette Cedex, France.
J Biol Chem. 1998 Aug 28;273(35):22480-9. doi: 10.1074/jbc.273.35.22480.
The changes in gene expression underlying the yeast adaptive stress response to H2O2 were analyzed by comparative two-dimensional gel electrophoresis of total cell proteins. The synthesis of at least 115 proteins is stimulated by H2O2, whereas 52 other proteins are repressed by this treatment. We have identified 71 of the stimulated and 44 of the repressed targets. The kinetics and dose-response parameters of the H2O2 genomic response were also analyzed. Identification of these proteins and their mapping into specific cellular processes give a distinct picture of the way in which yeast cells adapt to oxidative stress. As expected, H2O2-responsive targets include an important number of heat shock proteins and proteins with reactive oxygen intermediate scavenging activities. Exposure to H2O2 also results in a slowdown of protein biosynthetic processes and a stimulation of protein degradation pathways. Finally, the most remarkable result inferred from this study is the resetting of carbohydrate metabolism minutes after the exposure to H2O2. Carbohydrate fluxes are redirected to the regeneration of NADPH at the expense of glycolysis. This study represents the first genome-wide characterization of a H2O2-inducible stimulon in a eukaryote.
通过对全细胞蛋白质进行比较二维凝胶电泳,分析了酵母对过氧化氢适应性应激反应背后的基因表达变化。至少115种蛋白质的合成受到过氧化氢的刺激,而另外52种蛋白质则受到这种处理的抑制。我们已经鉴定出71种受刺激的靶标和44种受抑制的靶标。还分析了过氧化氢基因组反应的动力学和剂量反应参数。对这些蛋白质的鉴定以及将它们映射到特定细胞过程中,清晰地展现了酵母细胞适应氧化应激的方式。正如预期的那样,过氧化氢反应性靶标包括大量热休克蛋白和具有活性氧中间物清除活性的蛋白质。暴露于过氧化氢还会导致蛋白质生物合成过程放缓,并刺激蛋白质降解途径。最后,从这项研究中推断出的最显著结果是,在暴露于过氧化氢几分钟后,碳水化合物代谢发生了重置。碳水化合物通量被重新导向以糖酵解为代价的NADPH再生。这项研究代表了真核生物中过氧化氢诱导刺激子的首次全基因组表征。