Research Institute of Micro-Nano Technology, Hosei Universitygrid.257114.4, Koganei, Tokyo, Japan.
Department of Life Sciences, Prefectural University of Hiroshimagrid.412155.6, Shobara, Hiroshima, Japan.
J Bacteriol. 2022 Jan 18;204(1):e0042021. doi: 10.1128/JB.00420-21. Epub 2021 Oct 25.
Escherichia coli survives under acid stress conditions by the glutamic acid-dependent acid resistance (GAD) system, which enzymatically decreases intracellular protons. We found a linkage between GAD and flagellar systems in E. coli. The gene, one of the GAD cluster genes, encodes an uncharacterized membrane protein. A reporter assay showed that the promoter was induced in a GadE-dependent manner when grown in the M9 glycerol medium. Transcriptome analysis revealed that most of the transcripts were from genes involved in flagellum synthesis, and cell motility increased not only in the -deficient mutant but also in the -deficient mutant. Defects in both the and increased the intracellular level of FliA, an alternative sigma factor for flagellum synthesis, activated by the master regulator FlhDC. The promoter activity of the gene, which encodes repressor for the operon, was found to decrease in both the - and -deficient mutants. Transmission electron microscopy showed that the number of flagellar filaments on the -, -, and -deficient cells increased, and all three mutants showed higher motility than the parent strain. Thus, HdeD in the GAD system activates the promoter, resulting in a decrease in flagellar filaments in E. coli cells. We speculated that the synthesis of HdeD, stimulated in E. coli exposed to acid stress, could control the flagellum biosynthesis by sensing slight changes in pH at the cytoplasmic membrane. This could help in saving energy through termination of flagellum biosynthesis and improve bacterial survival efficiency within the animal digestive system. E. coli cells encounter various environments from the mouth down to the intestines within the host animals. The pH of gastric juice is lower than 2.0, and the bacterial must quickly respond and adapt to the following environmental changes before reaching the intestines. The quick response plays a role in cellular survival in the population, whereas adaptation may contribute to species survival. The GAD and flagellar systems are important for response to low pH in E. coli. Here, we identified the novel inner membrane regulator HdeD, encoding in the GAD cluster, to repress the synthesis of flagella. These insights provide a deeper understanding of how the bacteria enter the animal digestive system, survive, and form colonies in the intestines.
大肠杆菌通过谷氨酸依赖的酸抗性(GAD)系统在酸性应激条件下存活,该系统通过酶促作用降低细胞内的质子。我们发现大肠杆菌中的 GAD 系统与鞭毛系统之间存在联系。基因,GAD 簇基因之一,编码一种未被描述的膜蛋白。报告基因实验表明,在 M9 甘油培养基中生长时,以 GadE 依赖性方式诱导启动子。转录组分析显示,大多数转录本来自参与鞭毛合成的基因,并且不仅在缺陷突变体中,而且在缺陷突变体中细胞运动性都增加。和缺陷都增加了鞭毛合成的替代 sigma 因子 FliA 的细胞内水平,该因子由主调控因子 FlhDC 激活。发现编码 操纵子抑制剂的基因的启动子活性在和缺陷突变体中均降低。透射电子显微镜显示,鞭毛丝的数量在 -、- 和 - 缺陷细胞上增加,并且所有三个突变体的运动性均高于亲本菌株。因此,GAD 系统中的 HdeD 激活启动子,导致大肠杆菌细胞中的鞭毛丝减少。我们推测,暴露于酸性应激下的大肠杆菌中 HdeD 的合成可以通过感测细胞质膜上的 pH 细微变化来控制鞭毛生物合成。这有助于通过终止鞭毛生物合成来节省能量,并提高细菌在动物消化系统中的生存效率。大肠杆菌细胞在宿主动物体内从口腔到肠道遇到各种环境。胃液的 pH 值低于 2.0,细菌必须在到达肠道之前快速响应并适应以下环境变化。快速响应在细胞群中的细胞存活中发挥作用,而适应可能有助于物种存活。GAD 和鞭毛系统对大肠杆菌中低 pH 的响应很重要。在这里,我们鉴定了 GAD 簇中编码的新型内膜调节剂 HdeD,以抑制鞭毛的合成。这些见解提供了对细菌如何进入动物消化系统、在肠道中存活并形成菌落的更深入了解。