Bobrov Alexander G, Kirillina Olga, Fetherston Jacqueline D, Miller M Clarke, Burlison Joseph A, Perry Robert D
Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky, Lexington, KY, USA.
Mol Microbiol. 2014 Aug;93(4):759-75. doi: 10.1111/mmi.12693. Epub 2014 Jul 16.
Bacterial pathogens must overcome host sequestration of zinc (Zn(2+) ), an essential micronutrient, during the infectious disease process. While the mechanisms to acquire chelated Zn(2+) by bacteria are largely undefined, many pathogens rely upon the ZnuABC family of ABC transporters. Here we show that in Yersinia pestis, irp2, a gene encoding the synthetase (HMWP2) for the siderophore yersiniabactin (Ybt) is required for growth under Zn(2+) -deficient conditions in a strain lacking ZnuABC. Moreover, growth stimulation with exogenous, purified apo-Ybt provides evidence that Ybt may serve as a zincophore for Zn(2+) acquisition. Studies with the Zn(2+) -dependent transcriptional reporter znuA::lacZ indicate that the ability to synthesize Ybt affects the levels of intracellular Zn(2+) . However, the outer membrane receptor Psn and TonB as well as the inner membrane (IM) ABC transporter YbtPQ, which are required for Fe(3+) acquisition by Ybt, are not needed for Ybt-dependent Zn(2+) uptake. In contrast, the predicted IM protein YbtX, a member of the Major Facilitator Superfamily, was essential for Ybt-dependent Zn(2+) uptake. Finally, we show that the ZnuABC system and the Ybt synthetase HMWP2, presumably by Ybt synthesis, both contribute to the development of a lethal infection in a septicaemic plague mouse model.
在感染性疾病过程中,细菌病原体必须克服宿主对锌(Zn(2+))的螯合作用,锌是一种必需的微量营养素。虽然细菌获取螯合态Zn(2+)的机制在很大程度上尚不清楚,但许多病原体依赖于ABC转运蛋白的ZnuABC家族。在这里,我们表明,在鼠疫耶尔森菌中,在缺乏ZnuABC的菌株中,irp2基因(编码铁载体耶尔森菌素(Ybt)的合成酶(HMWP2))是在Zn(2+)缺乏条件下生长所必需的。此外,用外源性纯化的脱辅基Ybt进行生长刺激提供了证据,表明Ybt可能作为一种锌载体用于获取Zn(2+)。对依赖Zn(2+)的转录报告基因znuA::lacZ的研究表明,合成Ybt的能力会影响细胞内Zn(2+)的水平。然而,Ybt获取Fe(3+)所需的外膜受体Psn和TonB以及内膜(IM)ABC转运蛋白YbtPQ对于依赖Ybt的Zn(2+)摄取并不是必需的。相反,预测的内膜蛋白YbtX(主要促进剂超家族的成员)对于依赖Ybt的Zn(2+)摄取至关重要。最后,我们表明ZnuABC系统和Ybt合成酶HMWP2(可能通过Ybt合成)都有助于在败血性鼠疫小鼠模型中发展致死性感染。