Discovery of functional toxin/antitoxin systems in bacteria by shotgun cloning.

Discovery of functional toxin/antitoxin systems in bacteria by shotgun cloning.
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DOI:
10.1016/j.molcel.2013.02.002
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发表时间:
2013-04-11
期刊:
影响因子:
16
通讯作者:
Sorek, Rotem
Sorek, Rotem
中科院分区:
生物学1区
文献类型:
--
作者:
Sberro, Hila;Leavitt, Azita;Kiro, Ruth;Koh, Eugene;Peleg, Yoav;Qimron, Udi;Sorek, Rotem

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毒素-抗毒素(Toxin-antitoxin,TA)模块由一个毒性蛋白和一个对抗性抗毒素组成,在细菌生理学中起着重要作用。我们使用超过850万个随机克隆,研究了来自388个微生物基因组的150万个基因插入大肠杆菌宿主的实验。这揭示了数百个基因(毒素),只有当相邻基因(抗毒素)存在于同一克隆中时才能克隆。这些基因的聚类揭示了新的TA家族广泛存在于细菌基因组中,其中一些偏离了先前描述的这些模块的经典特征。将这些基因导入E.大肠杆菌的毒性试验表明,抗毒素可减轻毒素的毒性。T7噬菌体感染实验表明,两个新的模块可以提供针对噬菌体的抗性。此外,我们的实验揭示了噬菌体T7中的“抗防御”蛋白,其中和噬菌体抗性。我们的研究结果暴露了细菌和噬菌体之间军备竞赛的活跃前沿。
Toxin-antitoxin (TA) modules, composed of a toxic protein and a counteracting antitoxin, play important roles in bacterial physiology. We examined the experimental insertion of 1.5 million genes from 388 microbial genomes into an Escherichia coli host using over 8.5 million random clones. This revealed hundreds of genes (toxins) that could only be cloned when the neighboring gene (antitoxin) was present on the same clone. Clustering of these genes revealed novel TA families widespread in bacterial genomes, some of which deviate from the classical characteristics previously described for such modules. Introduction of these genes into E. coli validated that the toxin toxicity is mitigated by the antitoxin. Infection experiments with T7 phage showed that two of the new modules can provide resistance against phage. Moreover, our experiments revealed an 'anti-defense' protein in phage T7 that neutralizes phage resistance. Our results expose active fronts in the arms race between bacteria and phage.
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