Identification and characterization of a novel type II toxin-antitoxin system in Aeromonas veronii

IF 2.3 3区 生物学 Q3 MICROBIOLOGY
Caihong Ji, Ting He, Binbin Wu, Xiaomei Cao, Xiaping Fan, Xia Liu, Xiaodan Li, Miao Yang, Jihan Wang, Ling Xu, Shengbiao Hu, Liqiu Xia, Yunjun Sun
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Abstract

The bacterial type II toxin-antitoxin (TA) system is a rich genetic element that participates in various physiological processes. Aeromonas veronii is the main bacterial pathogen threatening the freshwater aquaculture industry. However, the distribution of type II TA system in A. veronii was seldom documented and its roles in the life activities of A. veronii were still unexplored. In this study, a novel type II TA system AvtA-AvtT was predicted in a fish pathogen Aeromonas veronii biovar sobria with multi-drug resistance using TADB 2.0. Through an Escherichia coli host killing and rescue assay, we demonstrated that AvtA and AvtT worked as a genuine TA system, and the predicted toxin AvtT actually functioned as an antitoxin while the predicted antitoxin AvtA actually functioned as a toxin. The binding ability of AvtA with AvtT proteins were confirmed by dot blotting analysis and co-immunoprecipitation assay. Furthermore, we found that the toxin and antitoxin labelled with fluorescent proteins were co-localized. In addition, it was found that the transcription of AvtAT bicistronic operon was repressed by the AvtAT protein complex. Deletion of avtA gene and avtT gene had no obvious effect on the drug susceptibility. This study provides first characterization of type II TA system AvtA-AvtT in aquatic pathogen A. veronii.

Abstract Image

识别并鉴定维龙单胞菌的新型 II 型毒素-抗毒素系统。
细菌 II 型毒素-抗毒素(TA)系统是一种参与各种生理过程的丰富遗传因子。veronii 气单胞菌是威胁淡水养殖业的主要细菌病原体。然而,Ⅱ型TA系统在蚯蚓气单胞菌中的分布很少被记录,其在蚯蚓气单胞菌生命活动中的作用也尚未被探索。本研究利用 TADB 2.0 预测了具有多重耐药性的鱼类病原体 Aeromonas veronii biovar sobria 中的新型 II 型 TA 系统 AvtA-AvtT。通过大肠杆菌宿主杀伤和拯救试验,我们证明了AvtA和AvtT是真正的TA系统,预测的毒素AvtT实际上起着抗毒素的作用,而预测的抗毒素AvtA实际上起着毒素的作用。通过点印迹分析和共免疫沉淀试验,我们证实了AvtA与AvtT蛋白的结合能力。此外,我们还发现用荧光蛋白标记的毒素和抗毒素是共定位的。此外,我们还发现 AvtAT 双核苷酸操作子的转录受到 AvtAT 蛋白复合物的抑制。删除 avtA 基因和 avtT 基因对药物敏感性无明显影响。本研究首次鉴定了水生病原体A. veronii的II型TA系统AvtA-AvtT。
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来源期刊
Archives of Microbiology
Archives of Microbiology 生物-微生物学
CiteScore
4.90
自引率
3.60%
发文量
601
审稿时长
3 months
期刊介绍: Research papers must make a significant and original contribution to microbiology and be of interest to a broad readership. The results of any experimental approach that meets these objectives are welcome, particularly biochemical, molecular genetic, physiological, and/or physical investigations into microbial cells and their interactions with their environments, including their eukaryotic hosts. Mini-reviews in areas of special topical interest and papers on medical microbiology, ecology and systematics, including description of novel taxa, are also published. Theoretical papers and those that report on the analysis or ''mining'' of data are acceptable in principle if new information, interpretations, or hypotheses emerge.
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