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首页> 外文期刊>Applied Microbiology >Genome-Wide Screening for Identification of Novel Toxin-Antitoxin Systems in Staphylococcus aureus
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Genome-Wide Screening for Identification of Novel Toxin-Antitoxin Systems in Staphylococcus aureus

机译：全基因组筛选，用于鉴定金黄色葡萄球菌中新型毒素-抗毒素系统

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Toxin-antitoxin (TA) systems consist of toxin-inhibiting diverse cellular functions (e.g., DNA replication, transcription, and translation) and a noncoding RNA or protein antitoxin. TA systems are associated with various cellular events, such as stress responses, programmed cell death, and bacterial pathogenicity. Recent advances in genome sequencing and bioinformatics research have demonstrated that most bacteria harbor various kinds of TA modules on their chromosomes; however, there is little understanding of chromosomally encoded TA systems in the Gram-positive pathogen Staphylococcus aureus. Here, we report on newly discovered S. aureus TA systems, each of which is composed of two proteins. Manual search and gene operon prediction analysis identified eight 2-gene operons as potential candidates for TA systems. Subsequently, using an Escherichia coli host killing and rescue assay, we demonstrated that four of the eight candidates worked as TA systems, designated tsaAT, tsbAT, tscAT, and tsdAT. Moreover, the TsaT, TsbT, TscT, and TsdT toxins inhibited S. aureus growth, and the toxicity of TsbT was neutralized by coexpressing the tsbA gene in the native host, S. aureus. Further, the bioinformatics analysis of the gene clusters revealed that TsaAT, TsbAT, TscAT, and TsdAT did not exhibit sequence similarity to known bacterial TA systems, and their homologues were present only within Staphylococcus species and not among any other bacteria. Our results further advance not only the understanding of S. aureus TA systems but also the study of unannotated TA systems in various bacterial species.IMPORTANCE Recent advances in genome sequencing and bioinformatics research have demonstrated that most pathogenic bacteria harbor a large number of chromosomally encoded toxin-antitoxin (TA) modules. However, little is known about the TA systems in S. aureus. Here, we newly identified four S. aureus TA systems using a combination of manual base-by-base screening and functional analysis in E. coli. Moreover, all toxins of the identified TA systems caused growth inhibition in the native host S. aureus. Although the newly identified TA systems did not exhibit sequence similarity with known bacterial TA systems, their orthologues were conserved only among other Staphylococcus species, indicating their uniqueness to staphylococci. Our approach opens the possibility for studying unannotated TA systems in various bacterial species.
机译：毒素-抗毒素（TA）系统由抑制毒素的多种细胞功能（例如DNA复制，转录和翻译）和非编码RNA或蛋白质抗毒素组成。 TA系统与各种细胞事件相关，例如应激反应，程序性细胞死亡和细菌致病性。基因组测序和生物信息学研究的最新进展表明，大多数细菌的染色体上都带有各种TA模块。然而，对革兰氏阳性病原体金黄色葡萄球菌中的染色体编码TA系统了解甚少。在这里，我们报告新发现的金黄色葡萄球菌TA系统，每个系统由两种蛋白质组成。手动搜索和基因操纵子预测分析确定了八个2基因操纵子作为TA系统的潜在候选者。随后，使用大肠杆菌宿主的杀伤和拯救实验，我们证明了八个候选者中的四个作为TA系统工作，分别命名为tsaAT，tsbAT，tscAT和tsdAT。此外，TsaT，TsbT，TscT和TsdT毒素抑制了金黄色葡萄球菌的生长，并且通过在天然宿主金黄色葡萄球菌中共表达tsbA基因来中和TsbT的毒性。此外，对基因簇的生物信息学分析表明，TsaAT，TsbAT，TscAT和TsdAT与已知的细菌TA系统没有序列相似性，它们的同源物仅存在于葡萄球菌中，而在其他细菌中则不存在。我们的研究结果不仅进一步提高了对金黄色葡萄球菌TA系统的理解，而且进一步促进了各种细菌物种中未注释的TA系统的研究。重要信息在基因组测序和生物信息学研究方面的最新进展表明，大多数致病细菌都具有大量染色体编码的毒素-抗毒素（TA）模块。然而，关于金黄色葡萄球菌的TA系统知之甚少。在这里，我们通过在大肠杆菌中进行手动逐个碱基筛选和功能分析相结合，新鉴定了四个金黄色葡萄球菌TA系统。而且，所鉴定的TA系统的所有毒素在天然宿主金黄色葡萄球菌中引起生长抑制。尽管新近鉴定的TA系统与已知的细菌TA系统没有序列相似性，但它们的直向同源物仅在其他葡萄球菌物种中保守，表明它们对葡萄球菌的独特性。我们的方法为研究各种细菌中未注释的TA系统提供了可能性。

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