Rothhardt Johannes E, Kröger Carsten, Broadley Steven P, Fuchs Thilo M
Lehrstuhl für Mikrobielle Ökologie, Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Wissenschaftszentrum Weihenstephan, Technische Universität München, Weihenstephaner Berg 3, 85354, Freising, Germany.
Mol Microbiol. 2014 Nov;94(3):700-12. doi: 10.1111/mmi.12788. Epub 2014 Oct 6.
In Salmonella enterica serovar Typhimurium (S. Typhimurium), the genomic island GEI4417/4436 is responsible for the utilization of myo-inositol (MI) as carbon and energy source. Here, we report the characterization of a novel, island-encoded positive autoregulator termed ReiD (STM4423) that is specific to certain S. enterica strains and Escherichia coli strain ED1a able to use MI. ReiD was essential for growth with this polyol and also contributed to S. Typhimurium proliferation in swine caecum content. Providing higher copy numbers of ReiD reduced the long lag phase of 2 days during growth of S. Typhimurium in MI medium by 50%. In a heterologous host, expression of ReiD activated the transcription from the promoter of iolE/iolG, whose products catalyse the initial two steps in MI degradation. Episomal expression of iolE/iolG1 rescued the otherwise zero growth phenotype of a reiD deletion mutant in MI medium. Gel mobility shift assays with purified ReiD demonstrated directed interaction of ReiD with its own promoter and that of iolE. The repressor IolR bound the reiD promoter, implying that reiD is part of the IolR regulon. Taken together, the regulator ReiD is a trigger to accelerate the switch from more easily accessible nutrients to MI utilization by S. Typhimurium.
在肠炎沙门氏菌鼠伤寒血清型(鼠伤寒沙门氏菌)中,基因组岛GEI4417/4436负责利用肌醇(MI)作为碳源和能源。在此,我们报告了一种新型的、由该岛编码的正向自调控因子ReiD(STM4423)的特性,它对某些肠炎沙门氏菌菌株和能够利用MI的大肠杆菌菌株ED1a具有特异性。ReiD对于利用这种多元醇进行生长至关重要,并且也有助于鼠伤寒沙门氏菌在猪盲肠内容物中增殖。提供更高拷贝数的ReiD可使鼠伤寒沙门氏菌在MI培养基中生长时2天的长延迟期缩短50%。在异源宿主中,ReiD的表达激活了iolE/iolG启动子的转录,其产物催化MI降解的最初两步。iolE/iolG1的附加型表达挽救了reiD缺失突变体在MI培养基中原本为零生长的表型。用纯化的ReiD进行凝胶迁移率变动分析表明,ReiD与其自身启动子以及iolE的启动子存在直接相互作用。阻遏物IolR结合reiD启动子,这意味着reiD是IolR调控子的一部分。综上所述,调控因子ReiD是促使鼠伤寒沙门氏菌从更容易利用的营养物质转向利用MI的一个触发因素。