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猪链球菌色氨酸转运蛋白 X 是色氨酸摄取系统的一部分,其表达受 T 盒调控元件调控。

Streptococcus suis TrpX is part of a tryptophan uptake system, and its expression is regulated by a T-box regulatory element.

机构信息

Institute for Microbiology, University of Veterinary Medicine Hannover, Hannover, Germany.

Unit of Microbiology and Immunology, Faculty of Veterinary, University of Zaragoza, Zaragoza, Spain.

出版信息

Sci Rep. 2022 Aug 17;12(1):13920. doi: 10.1038/s41598-022-18227-3.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b451/9385718/28652696994e/41598_2022_18227_Fig1_HTML.jpg

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