Institute for Microbiology, University of Veterinary Medicine Hannover, Hannover, Germany.
Unit of Microbiology and Immunology, Faculty of Veterinary, University of Zaragoza, Zaragoza, Spain.
Sci Rep. 2022 Aug 17;12(1):13920. doi: 10.1038/s41598-022-18227-3.
Streptococcus suis, a common member of the porcine respiratory microbiota, can cause life-threatening diseases in pigs as well as humans. A previous study identified the gene trpX as conditionally essential for in vivo survival by intrathecal infection of pigs with a transposon library of S. suis strain 10. Here, we characterized trpX, encoding a putative tryptophan/tyrosine transport system substrate-binding protein, in more detail. We compared growth capacities of the isogenic trpX-deficient mutant derivative strain 10∆trpX with its parent. Growth experiments in chemically defined media (CDM) revealed that growth of 10∆trpX depended on tryptophan concentration, suggesting TrpX involvement in tryptophan uptake. We demonstrated that trpX is part of an operon structure and co-transcribed with two additional genes encoding a putative permease and ATPase, respectively. Bioinformatics analysis identified a putative tryptophan T-box riboswitch in the 5' untranslated region of this operon. Finally, qRT-PCR and a reporter activation assay revealed trpX mRNA induction under tryptophan-limited conditions. In conclusion, our study showed that TrpX is part of a putative tryptophan ABC transporter system regulated by a T-box riboswitch probably functioning as a substrate-binding protein. Due to the tryptophan auxotrophy of S. suis, TrpX plays a crucial role for metabolic adaptation and growth during infection.
猪链球菌是猪呼吸道微生物群中的常见成员,它既能引起猪的致命性疾病,也能引起人类的致命性疾病。先前的一项研究通过对猪进行转座子文库的脑脊髓液感染,鉴定出基因 trpX 是体内生存的条件必需基因。在此,我们对编码假定色氨酸/酪氨酸运输系统底物结合蛋白的 trpX 进行了更详细的描述。我们比较了具有相同遗传背景的 trpX 缺陷突变体衍生株 10∆trpX 与其亲本的生长能力。在化学定义培养基(CDM)中的生长实验表明,10∆trpX 的生长依赖于色氨酸浓度,表明 TrpX 参与色氨酸摄取。我们证明 trpX 是操纵子结构的一部分,与另外两个分别编码假定透性酶和 ATP 酶的基因共转录。生物信息学分析在该操纵子的 5'非翻译区鉴定出一个假定的色氨酸 T -box 核糖开关。最后,qRT-PCR 和报告基因激活测定显示 trpX mRNA 在色氨酸限制条件下诱导。总之,我们的研究表明,TrpX 是假定色氨酸 ABC 转运系统的一部分,该系统受 T 盒核糖开关调控,可能作为一种底物结合蛋白发挥作用。由于猪链球菌的色氨酸营养缺陷,TrpX 在感染期间的代谢适应和生长中起着至关重要的作用。