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Vibrio cholerae VciB Promotes Iron Uptake via Ferrous Iron Transporters▿

机译:霍乱弧菌VciB通过亚铁转运蛋白促进铁吸收▿

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摘要

Vibrio cholerae uses a variety of strategies for obtaining iron in its diverse environments. In this study we report the identification of a novel iron utilization protein in V. cholerae, VciB. The vciB gene and its linked gene, vciA, were isolated in a screen for V. cholerae genes that permitted growth of an Escherichia coli siderophore mutant in low-iron medium. The vciAB operon encodes a predicted TonB-dependent outer membrane receptor, VciA, and a putative inner membrane protein, VciB. VciB, but not VciA, was required for growth stimulation of E. coli and Shigella flexneri strains in low-iron medium. Consistent with these findings, TonB was not needed for VciB-mediated growth. No growth enhancement was seen when vciB was expressed in an E. coli or S. flexneri strain defective for the ferrous iron transporter Feo. Supplying the E. coli feo mutant with a plasmid encoding either E. coli or V. cholerae Feo, or the S. flexneri ferrous iron transport system Sit, restored VciB-mediated growth; however, no stimulation was seen when either of the ferric uptake systems V. cholerae Fbp and Haemophilus influenzae Hit was expressed. These data indicate that VciB functions by promoting iron uptake via a ferrous, but not ferric, iron transport system. VciB-dependent iron accumulation via Feo was demonstrated directly in iron transport assays using radiolabeled iron. A V. cholerae vciB mutant did not exhibit any growth defects in either in vitro or in vivo assays, possibly due to the presence of other systems with overlapping functions in this pathogen.
机译:霍乱弧菌使用多种策略在其不同的环境中获取铁。在这项研究中,我们报告了霍乱弧菌VciB中一种新型铁利用蛋白的鉴定。在针对霍乱弧菌基因的筛选中分离了vciB基因及其连接的基因vciA,该基因允许在低铁培养基中生长大肠杆菌铁载体突变体。 vciAB操纵子编码一个预测的TonB依赖性外膜受体VciA和一个假定的内膜蛋白VciB。在低铁培养基中刺激大肠杆菌和弗氏志贺氏菌菌株的生长需要VciB,而不是VciA。与这些发现一致,VciB介导的生长不需要TonB。当vciB在有缺陷的亚铁转运蛋白Feo的大肠杆菌或弗氏链球菌中表达时,未见生长增强。向大肠杆菌feo突变体提供编码大肠杆菌或霍乱弧菌Feo或弗氏链球菌亚铁转运系统Sit的质粒,可恢复VciB介导的生长。然而,当表达铁的霍乱弧菌Fbp和流感嗜血杆菌Hit时,均未见刺激。这些数据表明,VciB通过促进通过铁(而非铁)铁运输系统吸收铁而起作用。通过放射性标记的铁直接在铁转运测定中证明了通过Feo产生的VciB依赖性铁积累。霍乱弧菌vciB突变体在体外或体内试验中均未显示任何生长缺陷,这可能是由于该病原体中存在其他具有重叠功能的系统所致。

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