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首页> 美国卫生研究院文献>Microbiology >Escherichia coli enterobactin synthesis and uptake mutants are hypersensitive to an antimicrobial peptide that limits the availability of iron in addition to blocking Holliday junction resolution
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Escherichia coli enterobactin synthesis and uptake mutants are hypersensitive to an antimicrobial peptide that limits the availability of iron in addition to blocking Holliday junction resolution

机译：大肠杆菌肠杆菌素合成和摄取突变体对抗菌肽过敏该抗菌肽除了阻止霍利迪结的解析外还限制了铁的可用性

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摘要

The peptide wrwycr inhibits Holliday junction resolution and is a potent antimicrobial. To study the physiological effects of wrwycr treatment on Escherichia coli cells, we partially screened the Keio collection of knockout mutants for those with increased sensitivity to wrwycr. Strains lacking part of the ferric-enterobactin (iron-bound siderophore) uptake and utilization system, parts of the enterobactin synthesis pathway, TolC (an outer-membrane channel protein) or Fur (an iron-responsive regulator) were hypersensitive to wrwycr. We provide evidence that the ΔtolC mutant was hypersensitive to wrwycr due to its reduced ability to efflux wrwycr from the cell rather than due to its export of newly synthesized enterobactin. Deleting ryhB, which encodes a small RNA involved in iron regulation, mostly relieved the wrwycr hypersensitivity of the fur and ferric-enterobactin uptake mutants, indicating that the altered regulation of a RyhB-controlled gene was at least partly responsible for the hypersensitivity of these strains. Chelatable iron in the cell, measured by electron paramagnetic resonance spectroscopy, increased dramatically following wrwycr treatment, as did expression of Fur-repressed genes and, to some extent, mutation frequency. These incongruous results suggest that while wrwycr treatment caused accumulation of chelatable iron in the cell, iron was not available to bind to Fur. This is corroborated by the observed induction of the suf system, which assembles iron–sulfur clusters in low-iron conditions. Disruption of iron metabolism by wrwycr, in addition to its effects on DNA repair, may make it a particularly effective antimicrobial in the context of the low-iron environment of a mammalian host.
机译：wrwycr肽抑制霍利迪结的分离，是一种有效的抗菌剂。为了研究wrwycr处理对大肠杆菌细胞的生理影响，我们部分筛选了Keio基因敲除突变体，以寻找对wrwycr敏感的突变体。缺乏部分铁-肠杆菌素（铁结合铁载体）的吸收和利用系统，部分肠杆菌素合成途径，TolC（外膜通道蛋白）或Fur（一种铁反应性调节剂）对wrwycr过敏。我们提供的证据表明，ΔtolC突变体对wrwycr高度敏感，这是由于其从细胞外排wrwycr的能力降低，而不是由于其新合成的肠杆菌素的输出。删除编码涉及铁调节的小RNA的ryhB，可以大部分缓解皮草和铁-肠杆菌素摄取突变体的wrwycr超敏性，这表明RyhB控制基因的调节改变至少部分负责这些菌株的超敏性。 。通过电子顺磁共振波谱测量，细胞中的螯合铁在wrwycr处理后急剧增加，Fur抑制基因的表达也有所提高，并且在一定程度上突变频率也是如此。这些不一致的结果表明，尽管wrwycr处理导致细胞中可螯合的铁积累，但铁无法与Fur结合。观察到的suf系统的感应证实了这一点，该系统在低铁条件下组装了铁硫簇。在哺乳动物宿主的低铁环境下，wrwycr破坏铁代谢，除了对DNA修复产生影响外，还可能使其成为一种特别有效的抗菌剂。

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