Systematic genetic dissection of PTS in Vibrio cholerae uncovers a novel glucose transporter and a limited role for PTS during infection of a mammalian host.
Systematic genetic dissection of PTS in Vibrio cholerae uncovers a novel glucose transporter and a limited role for PTS during infection of a mammalian host.
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DOI：
10.1111/mmi.13646
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发表时间：
2017-05
影响因子：
3.6
通讯作者：
Dalia AB
中科院分区：
文献类型：
--
作者：
Hayes CA;Dalia TN;Dalia AB
微生物物种中高亲和力碳水化合物摄取的常见机制是磷酸烯醇化酸依赖性磷酸转移酶系统（PTS）。该系统包括一个共享的组件，EI，这是所有PTS运输所需的，和许多碳水化合物摄取转运蛋白。在霍乱弧菌中，有13种不同的PTS转运蛋白。由于该系统内的遗传冗余，这些转运蛋白中的每一种的碳水化合物特异性目前尚未定义。在这里，使用自然转化的多重基因组编辑（MuGENT），我们系统地剖析了霍乱弧菌中的PTS转运。具体地说，我们产生了一个突变株，缺乏所有13个PTS转运蛋白，并从这个菌株，我们创建了一个小组的突变体，其中每个表达一个单一的转运蛋白。使用这个面板，我们在很大程度上定义了每个PTS转运蛋白的碳水化合物特异性。此外，这项分析发现了一种新的葡萄糖转运蛋白。我们进一步定义了这种转运蛋白的机制，并表征了其调节。使用我们的13个PTS转运蛋白突变体，我们还提供了第一个明确的证据表明，PTS的碳水化合物转运是不必要的，在感染过程中的幼鼠模型的霍乱。总之，这项研究显示了多重基因组编辑如何用于快速剖析复杂的生物系统和微生物系统中的遗传冗余。
A common mechanism for high affinity carbohydrate uptake in microbial species is the phosphoenolpyruvate-dependent phosphotransferase system (PTS). This system consists of a shared component, EI, which is required for all PTS transport, and numerous carbohydrate uptake transporters. In V. cholerae, there are 13 distinct PTS transporters. Due to genetic redundancy within this system, the carbohydrate specificity of each of these transporters is not currently defined. Here, using multiplex genome editing by natural transformation (MuGENT), we systematically dissect PTS transport in V. cholerae. Specifically, we generated a mutant strain that lacks all 13 PTS transporters, and from this strain, we created a panel of mutants where each expresses a single transporter. Using this panel, we have largely defined the carbohydrate specificities of each PTS transporter. Additionally, this analysis uncovered a novel glucose transporter. We have further defined the mechanism of this transporter and characterized its regulation. Using our 13 PTS transporter mutant, we also provide the first clear evidence that carbohydrate transport by the PTS is not essential during infection in an infant mouse model of cholera. In summary, this study shows how multiplex genome editing can be used to rapidly dissect complex biological systems and genetic redundancy in microbial systems.