Department of Microbiology and Immunology, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, USA.
Mol Microbiol. 2020 Aug;114(2):185-199. doi: 10.1111/mmi.14505. Epub 2020 Apr 19.
Proteus mirabilis is a Gram-negative uropathogen and frequent cause of catheter-associated urinary tract infection (CAUTI). One important virulence factor is its urease enzyme, which requires nickel to be catalytically active. It is, therefore, hypothesized that nickel import is critical for P. mirabilis urease activity and pathogenesis during infection. P. mirabilis strain HI4320 encodes two putative nickel import systems, designated Nik and Ynt. By disrupting the substrate-binding proteins from each import system (nikA and yntA), we show that Ynt is the primary nickel importer, while Nik only compensates for loss of Ynt at high nickel concentrations. We further demonstrate that these are the only binding proteins capable of importing nickel for incorporation into the urease enzyme. Loss of either nickel-binding protein results in a significant fitness defect in a murine model of CAUTI, but YntA is more crucial as the yntA mutant was significantly outcompeted by the nikA mutant. Furthermore, despite the importance of nickel transport for hydrogenase activity, the sole contribution of yntA and nikA to virulence is due to their role in urease activity, as neither mutant exhibited a fitness defect when disrupted in a urease-negative background.
奇异变形杆菌是一种革兰氏阴性尿路病原体,也是导管相关性尿路感染(CAUTI)的常见病因。其一个重要的毒力因子是其脲酶酶,该酶需要镍才能具有催化活性。因此,人们假设镍的摄取对于奇异变形杆菌脲酶活性和感染期间的发病机制至关重要。奇异变形杆菌 HI4320 株编码两个假定的镍导入系统,分别命名为 Nik 和 Ynt。通过破坏每个导入系统(nikA 和 yntA)的底物结合蛋白,我们表明 Ynt 是主要的镍导入体,而 Nik 仅在高镍浓度下补偿 Ynt 的缺失。我们进一步证明,这些是唯一能够将镍导入用于脲酶酶的结合蛋白。失去任何一种镍结合蛋白都会导致 CAUTI 小鼠模型中的显著适应性缺陷,但 YntA 更为关键,因为 yntA 突变体在镍浓度较高时被 nikA 突变体显著竞争淘汰。此外,尽管镍转运对于氢化酶活性很重要,但 yntA 和 nikA 对毒力的唯一贡献是由于它们在脲酶活性中的作用,因为当在脲酶阴性背景中破坏时,这两个突变体都没有表现出适应性缺陷。