Bernhardt Jörg, Weibezahn Jimena, Scharf Christian, Hecker Michael
Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, 17487 Greifswald, Germany.
Genome Res. 2003 Feb;13(2):224-37. doi: 10.1101/gr.905003.
Dual channel imaging and warping of two-dimensional (2D) protein gels were used to visualize global changes of the gene expression patterns in growing Bacillus subtilis cells during entry into the stationary phase as triggered by glucose exhaustion. The 2D gels only depict single moments during the cells' growth cycle, but a sequential series of overlays obtained at specific points of the growth curve facilitates visualization of the developmental processes at the proteomics scale. During glucose starvation a substantial reprogramming of the protein synthesis pattern was found, with 150 proteins synthesized de novo and cessation of the synthesis of almost 400 proteins. Proteins induced following glucose starvation belong to two main regulation groups: general stress/starvation responses induced by different stresses or starvation stimuli (sigma(B)-dependent general stress regulon, stringent response, sporulation), and glucose-starvation-specific responses (drop in glycolysis, utilization of alternative carbon sources, gluconeogenesis). Using the dual channel approach, it was not only possible to identify those regulons or stimulons, but also to follow the fate of each single protein by the three-color code: red, newly induced but not yet accumulated; yellow, synthesized and accumulated; and green, still present, but no longer being synthesized. These green proteins, which represent a substantial part of the protein pool in the nongrowing cell, are not accessible by using DNA arrays. The combination of 2D gel electrophoresis and MALDI TOF mass spectrometry with the dual channel imaging technique provides a new and comprehensive view of the physiology of growing or starving bacterial cell populations, here for the case of the glucose-starvation response.
利用双通道成像和二维(2D)蛋白质凝胶变形技术,可视化了枯草芽孢杆菌在因葡萄糖耗尽进入稳定期过程中基因表达模式的整体变化。二维凝胶仅描绘了细胞生长周期中的单个时刻,但在生长曲线特定点获得的一系列连续叠加图有助于在蛋白质组学规模上可视化发育过程。在葡萄糖饥饿期间,发现蛋白质合成模式发生了重大重编程,有150种蛋白质重新合成,近400种蛋白质停止合成。葡萄糖饥饿后诱导的蛋白质属于两个主要调控组:由不同应激或饥饿刺激诱导的一般应激/饥饿反应(依赖σ(B)的一般应激调节子、严格反应、孢子形成),以及葡萄糖饥饿特异性反应(糖酵解下降、替代碳源的利用、糖异生)。使用双通道方法,不仅可以识别那些调节子或刺激子,还可以通过三色编码追踪每种单一蛋白质的命运:红色,新诱导但尚未积累;黄色,已合成并积累;绿色,仍然存在但不再合成。这些绿色蛋白质代表了非生长细胞中蛋白质库的很大一部分,使用DNA阵列无法检测到。二维凝胶电泳和基质辅助激光解吸电离飞行时间质谱与双通道成像技术的结合,为生长或饥饿细菌群体的生理学提供了一个新的全面视角,这里以葡萄糖饥饿反应为例。