Mey Alexandra R, Wyckoff Elizabeth E, Hoover Lindsey A, Fisher Carolyn R, Payne Shelley M
The University of Texas at Austin, Section of Molecular Genetics and Microbiology, Austin, TX 78712-1095, USA.
J Bacteriol. 2008 Sep;190(17):5953-62. doi: 10.1128/JB.00569-08. Epub 2008 Jun 27.
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.
霍乱弧菌在其多样的生存环境中采用多种策略获取铁元素。在本研究中,我们报告了在霍乱弧菌中鉴定出一种新型铁利用蛋白VciB。vciB基因及其连锁基因vciA是在筛选允许大肠杆菌铁载体突变体在低铁培养基中生长的霍乱弧菌基因时分离得到的。vciAB操纵子编码一个预测的依赖TonB的外膜受体VciA和一个假定的内膜蛋白VciB。在低铁培养基中,刺激大肠杆菌和福氏志贺氏菌菌株生长需要VciB而非VciA。与这些发现一致,VciB介导的生长不需要TonB。当vciB在亚铁转运蛋白Feo缺陷的大肠杆菌或福氏志贺氏菌菌株中表达时,未见生长增强。为大肠杆菌feo突变体提供编码大肠杆菌或霍乱弧菌Feo的质粒,或福氏志贺氏菌亚铁转运系统Sit,可恢复VciB介导的生长;然而,当表达霍乱弧菌铁摄取系统Fbp和流感嗜血杆菌Hit中的任何一个时,未见刺激作用。这些数据表明,VciB通过促进经由亚铁而非高铁铁转运系统的铁摄取发挥作用。在使用放射性标记铁的铁转运试验中直接证明了通过Feo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表现出任何生长缺陷,这可能是由于该病原体中存在其他功能重叠的系统。