Iwadate Yumi, Kato Jun-Ichi
Department of Biological Sciences, Graduate Schools of Science and Engineering, Tokyo Metropolitan University, Tokyo 192-0397, Japan.
Microbiology (Reading). 2017 Dec;163(12):1912-1923. doi: 10.1099/mic.0.000534. Epub 2017 Nov 17.
The Escherichia coli PhoB-PhoR two-component system responds to phosphate starvation and induces the expression of many genes. Previous studies suggested that phosphate starvation induces oxidative stress, but the involvement of the PhoB regulon in oxidative stress tolerance has not been clarified. Here, we showed that ytfK, one of the PhoB regulon genes, is involved in cell tolerance to a redox-cycling drug, menadione, and H2O2 in stationary-phase cells. A ytfK deletion mutant was sensitive to H2O2 when the cells were grown anaerobically or micro-aerobically in the presence of nitrate. Genetic analysis suggested that the ytfK gene has a functional relationship with the oxyR and fur genes, among the oxyR regulon, at least, a catalase-encoding katG gene and peroxidase-encoding ahpCF genes. Overproduction of YtfK resulted in a KatG-dependent decrease of H2O2 concentration in the cell suspension, suggesting that katG is one of the targets of YtfK. Using a katG'-lacZ reporter fusion, we showed that YtfK enhances the transcription of katG although it was not clarified whether YtfK functions directly or not. We also showed that ytfK disruption results in reduced viability of stationary-phase cells under phosphate starvation. These results indicated that YtfK is involved in H2O2 tolerance by stimulating directly or indirectly the transcription of at least the catalase gene, and that this system plays an important role in cellular survival during phosphate starvation.
大肠杆菌PhoB-PhoR双组分系统对磷酸盐饥饿作出反应并诱导许多基因的表达。先前的研究表明,磷酸盐饥饿会诱导氧化应激,但PhoB调控子在氧化应激耐受性中的作用尚未阐明。在此,我们表明,PhoB调控子基因之一ytfK参与了静止期细胞对氧化还原循环药物甲萘醌和H2O2的耐受性。当细胞在硝酸盐存在下厌氧或微需氧生长时,ytfK缺失突变体对H2O2敏感。遗传分析表明,ytfK基因与oxyR调控子中的oxyR和fur基因至少有功能关系,其中包括编码过氧化氢酶的katG基因和编码过氧化物酶的ahpCF基因。YtfK的过量表达导致细胞悬液中H2O2浓度以KatG依赖的方式降低,这表明katG是YtfK的靶标之一。使用katG'-lacZ报告基因融合,我们表明YtfK增强了katG的转录,尽管尚不清楚YtfK是否直接发挥作用。我们还表明,ytfK的破坏导致磷酸盐饥饿条件下静止期细胞的活力降低。这些结果表明,YtfK通过直接或间接刺激至少过氧化氢酶基因的转录而参与H2O2耐受性,并且该系统在磷酸盐饥饿期间的细胞存活中起重要作用。