Marouga R, Kjelleberg S
Department of General and Marine Microbiology, Göteborg University, Sweden.
J Bacteriol. 1996 Feb;178(3):817-22. doi: 10.1128/jb.178.3.817-822.1996.
Proteins induced during the initial phase of recovery after long-term carbon starvation in the marine Vibrio sp. strain S14 were identified by two-dimensional gel electrophoresis analysis. Nutritional upshift experiments with pulse-labeled cells were performed after addition of glucose to cells starved for 48 h. Eighteen proteins synthesized during the first 3 min after substrate addition were identified and designated immediate upshift proteins (Iup proteins). They were induced at least 10-fold compared with the rate of synthesis during starvation. Of the Iup proteins, five are not found in exponentially growing cells. Subsequent to the first 3 min of glucose addition, a complex pattern of sequential synthesis of proteins made during a transient phase as well as proteins made during 60 min of the outgrowth response was monitored. To resolve whether the Iup proteins were synthesized from stable transcripts, the initiation of transcription was inhibited by rifampin (Rif). Addition of Rif 5 min prior to glucose promoted upshift resulted in the synthesis of 12 Iup proteins. Furthermore, three Iup proteins were still induced by cells that were Rif treated 20 min prior to the upshift. These results suggest that stable but silent transcripts exist during starvation and that the translation of these mRNA species is initiated by substrate addition. This regulatory mechanism may be essential for an immediate initiation of the recovery program by the nongrowing cell.
通过二维凝胶电泳分析,鉴定了海洋弧菌属菌株S14在长期碳饥饿后恢复初始阶段诱导产生的蛋白质。在向饥饿48小时的细胞中添加葡萄糖后,对脉冲标记的细胞进行了营养上调实验。鉴定并命名了底物添加后最初3分钟内合成的18种蛋白质为即时上调蛋白(Iup蛋白)。与饥饿期间的合成速率相比,它们的诱导倍数至少为10倍。在Iup蛋白中,有5种在指数生长的细胞中未发现。在添加葡萄糖的最初3分钟之后,监测了在短暂阶段合成的蛋白质以及在生长反应60分钟内合成的蛋白质的复杂顺序合成模式。为了确定Iup蛋白是否由稳定的转录本合成,用利福平(Rif)抑制转录起始。在葡萄糖促进上调前5分钟添加Rif导致12种Iup蛋白的合成。此外,在上调前20分钟用Rif处理的细胞仍诱导产生三种Iup蛋白。这些结果表明,饥饿期间存在稳定但沉默的转录本,并且这些mRNA种类的翻译通过底物添加启动。这种调节机制对于非生长细胞立即启动恢复程序可能至关重要。