Tkachenko A G, Shumkov M S, Akhova A V
Institute of Ecology and Genetics of Microorganisms, Ural Branch of the Russian Academy of Sciences, 614081 Perm, Russia.
Biochemistry (Mosc). 2006 Feb;71(2):185-93. doi: 10.1134/s0006297906020118.
Metabolites accumulated in the culture medium of Escherichia coli cells induce expression of the rpoS gene encoding the alternative sigmaS subunit of RNA polymerase, which controls adaptation of E. coli to acid stress during growth in glucose-mineral medium. The effect of acetate and succinate as end products of E. coli metabolism has been investigated on the levels of transcription, translation, and sigmaS protein stability. These end products mainly influenced the stability of the RNA polymerase sigmaS subunit. Under conditions of acid stress caused by acetate addition, the content of polyamines in the cells and medium decreased, whereas artificial rpoS gene switch-off by antisense RNA was accompanied by increase in polyamine level. Addition of polyamine to E. coli cells treated with acetate and especially with succinate caused a significant concentration-dependent stimulatory effect on rpoS expression. Thus, induction of the rpoS regulon depends on the combined action of the investigated metabolites determining adequate control of gene expression under conditions of acid stress. A scheme for metabolic pathways describing the role of putrescine in the maintenance of intracellular pH and polyamine pool homeostasis during E. coli adaptation to acid stress is proposed.
在大肠杆菌细胞培养基中积累的代谢产物可诱导编码RNA聚合酶替代西格玛S亚基的rpoS基因表达,该亚基控制大肠杆菌在葡萄糖 - 矿物质培养基中生长期间对酸胁迫的适应性。已研究了作为大肠杆菌代谢终产物的乙酸盐和琥珀酸盐对转录、翻译水平以及西格玛S蛋白稳定性的影响。这些终产物主要影响RNA聚合酶西格玛S亚基的稳定性。在添加乙酸盐引起的酸胁迫条件下,细胞和培养基中的多胺含量降低,而通过反义RNA人为关闭rpoS基因则伴随着多胺水平的升高。向用乙酸盐尤其是琥珀酸盐处理的大肠杆菌细胞中添加多胺对rpoS表达产生了显著的浓度依赖性刺激作用。因此,rpoS调节子的诱导取决于所研究代谢产物的联合作用,从而在酸胁迫条件下对基因表达进行适当控制。提出了一个代谢途径方案,描述了腐胺在大肠杆菌适应酸胁迫过程中维持细胞内pH和多胺库稳态的作用。