Siarot Lowela, Toyazaki Hiroki, Hidaka Makoto, Kurumisawa Keigo, Hirakawa Tomoki, Morohashi Kengo, Aono Toshihiro
Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
Appl Environ Microbiol. 2017 Sep 15;83(19). doi: 10.1128/AEM.01197-17. Print 2017 Oct 1.
Bacteria have multiple K uptake systems. , for example, has three types of K uptake systems, which include the low-K-inducible KdpFABC system and two constitutive systems, Trk (TrkAG and TrkAH) and Kup. ORS571, a rhizobium that forms nitrogen-fixing nodules on the stems and roots of , also has three types of K uptake systems. Through phylogenetic analysis, we found that has two genes homologous to and , designated and We also found that is adjacent to in the genome and these two genes are transcribed as an operon; however, is present at a distinct locus. Our results demonstrated that , , and were expressed in the wild-type stem nodules, whereas was not. Interestingly, Δ and Δ Δ mutants formed Fix nodules, while the Δ Δ Δ Δ mutant formed Fix nodules, suggesting that with the additional deletion of Trk system genes in the Δ mutant, Fix nodule phenotypes were recovered. of the Δ Δ mutant was expressed in stem nodules, but not in the free-living state, under high-K conditions. However, of the Δ Δ Δ Δ mutant was highly expressed even under high-K conditions. The cytoplasmic K levels in the Δ Δ Δ mutant, which did not express under high-K conditions, were markedly lower than those in the Δ Δ Δ Δ mutant. Taking all these results into consideration, we propose that TrkJ is involved in the repression of in response to high external K concentrations and that the TrkAI system is unable to function in stem nodules. K is a major cytoplasmic cation in prokaryotic and eukaryotic cells. Bacteria have multiple K uptake systems to control the cytoplasmic K levels. In many bacteria, the K uptake system KdpFABC is expressed under low-K conditions. For years, many researchers have argued over how bacteria sense K concentrations. Although KdpD of is known to sense both cytoplasmic and extracellular K concentrations, the detailed mechanism of K sensing is still unclear. In this study, we propose that the transmembrane TrkJ protein of acts as a sensor for the extracellular K concentration and that high extracellular K concentrations repress the expression of KdpFABC via TrkJ.
细菌有多种钾摄取系统。例如,大肠杆菌有三种类型的钾摄取系统,包括低钾诱导型KdpFABC系统和两种组成型系统Trk(TrkAG和TrkAH)以及Kup。ORS571是一种能在百脉根茎和根上形成固氮根瘤的根瘤菌,它也有三种类型的钾摄取系统。通过系统发育分析,我们发现百脉根有两个与大肠杆菌的kdpD和kdpE同源的基因,分别命名为LjkdpD和LjkdpE。我们还发现LjkdpD在基因组中与LjkdpE相邻,这两个基因作为一个操纵子转录;然而,Ljkup位于一个不同的位点。我们的结果表明,LjkdpD、LjkdpE和Ljkup在野生型茎根瘤中表达,而LjtrkA不表达。有趣的是,ΔLjkdpD和ΔLjkdpEΔLjkup突变体形成Fix根瘤,而ΔLjtrkAΔLjtrkBΔLjtrkDΔLjtrkG突变体形成Fix根瘤,这表明在ΔLjtrkA突变体中额外缺失Trk系统基因后,Fix根瘤表型得以恢复。ΔLjkdpDΔLjkdpE突变体的Ljkup在茎根瘤中表达,但在高钾条件下的自由生活状态下不表达。然而,ΔLjtrkAΔLjtrkBΔLjtrkDΔLjtrkG突变体的Ljkup即使在高钾条件下也高度表达。在高钾条件下不表达Ljkup的ΔLjkdpDΔLjkdpE突变体的细胞质钾水平明显低于ΔLjtrkAΔLjtrkBΔLjtrkDΔLjtrkG突变体。综合所有这些结果,我们提出TrkJ参与了对高细胞外钾浓度下Ljkup表达的抑制,并且TrkAI系统在茎根瘤中无法发挥作用。钾是原核和真核细胞中的主要细胞质阳离子。细菌有多种钾摄取系统来控制细胞质钾水平。在许多细菌中,钾摄取系统KdpFABC在低钾条件下表达。多年来,许多研究人员一直在争论细菌如何感知钾浓度。尽管已知大肠杆菌的KdpD能感知细胞质和细胞外钾浓度,但钾感知的详细机制仍不清楚。在这项研究中,我们提出百脉根的跨膜TrkJ蛋白作为细胞外钾浓度的传感器,高细胞外钾浓度通过TrkJ抑制KdpFABC的表达。