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Characterization of the Yersinia pestisYfu ABC Inorganic Iron Transport System

机译：鼠疫耶尔森菌Yfu ABC无机铁运输系统的表征

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In Yersinia pestis, the causative agent of plague, two inorganic iron transport systems have been partially characterized. The yersiniabactin (Ybt) system is a siderophore-dependent transport system required for full virulence. Yfe is an ABC transport system that accumulates both iron and manganese. We have identified and cloned aY. pestis yfuABC operon. The YfuABC system is a member of the cluster of bacterial ABC iron transporters that include Sfu ofSerratia, Hit of Haemophilus, and Yfu ofYersinia enterocolitica. The Y. pestis KIM6+ system is most homologous to that in Y. enterocolitica, showing identities of 84% for YfuA (periplasmic binding protein), 87% for YfuB (inner membrane permease), and 75% for YfuC (ATP hydrolase). We constructed a yfuABC promoter-lacZ fusion to examine regulation of transcription. This promoter contains a potential Fur binding sequence and is iron and Fur regulated. Significant expression from the yfuABC promoter occurred during iron-deficient growth conditions. In vitro transcription and translation of a recombinant plasmid encoding yfuABCindicates that YfuABC proteins are expressed. Escherichia coli 1017 (an enterobactin-deficient mutant) carrying this plasmid was able to grow in an iron-restrictive complex medium. We constructed a deletion encompassing the yfuABC promoter and most of yfuA. This mutation was introduced into strains with mutations in Ybt, Yfe, or both systems to examine the role of Yfu in iron acquisition in Y. pestis. Growth of theyfu mutants in a deferrated, defined medium (PMH2) at 26 and 37°C failed to identify a growth or iron transport defect due to the yfu mutation. Fifty percent lethal dose studies in mice did not demonstrate a role for the Yfu system in mammalian virulence.
机译：在鼠疫耶尔森菌中，鼠疫的病原体部分表征了两个无机铁的运输系统。耶尔西菌素（Yersiniabactin，Ybt）系统是完全毒力所需的铁载体依赖性转运系统。 Yfe是一种ABC传输系统，可以同时收集铁和锰。我们已经识别并克隆了一个 Y。鼠疫yfuABC 操纵子。 YfuABC系统是细菌ABC铁转运蛋白簇的成员，包括 Serratia 的Sfu，嗜血杆菌的命中和 Enterocolitica 的Yfu。 。 Y。鼠疫 KIM6 +系统与 Y中的同源性最高。肠结肠炎显示，YfuA（周质结合蛋白）的身份为84％，YfuB（内膜渗透酶）的身份为87％，YfuC（ATP水解酶）的身份为75％。我们构建了一个 yfuABC 启动子- lacZ 融合体来研究转录调控。该启动子含有潜在的Fur结合序列，并且受铁和Fur调节。在缺铁的生长条件下， yfuABC 启动子显着表达。编码 yfuABC 的重组质粒的体外转录和翻译表明表达了YfuABC蛋白。携带该质粒的大肠杆菌 1017（肠杆菌素缺陷型突变体）能够在铁限制型复合培养基中生长。我们构建了一个包含 yfuABC 启动子和大部分 yfuA 的缺失。将此突变引入Ybt，Yfe或两个系统均具有突变的菌株中，以检验Yfu在Y中铁的获取中的作用。瘟疫。在 yfu 突变体在26°C和37°C的延缓，确定培养基（PMH2）中的生长未能识别出由于 yfu 突变引起的生长或铁转运缺陷。小鼠中百分之五十的致死剂量研究并未证明Yfu系统在哺乳动物毒力中的作用。

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