Riboldi Gustavo Pelicioli, Bierhals Christine Garcia, Mattos Eduardo Preusser de, Frazzon Ana Paula Guedes, d'Azevedo Pedro Alves, Frazzon Jeverson
Laboratório de Cocos Gram-positivos e Microbiologia Molecular, Departamento de Microbiologia, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, RS, Brasil.
Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil.
Mem Inst Oswaldo Cruz. 2014 Jul;109(4):408-13. doi: 10.1590/0074-0276140006. Epub 2014 Jun 6.
The Firmicutes bacteria participate extensively in virulence and pathological processes. Enterococcus faecalis is a commensal microorganism; however, it is also a pathogenic bacterium mainly associated with nosocomial infections in immunocompromised patients. Iron-sulfur [Fe-S] clusters are inorganic prosthetic groups involved in diverse biological processes, whose in vivo formation requires several specific protein machineries. Escherichia coli is one of the most frequently studied microorganisms regarding [Fe-S] cluster biogenesis and encodes the iron-sulfur cluster and sulfur assimilation systems. In Firmicutes species, a unique operon composed of the sufCDSUB genes is responsible for [Fe-S] cluster biogenesis. The aim of this study was to investigate the potential of the E. faecalis sufCDSUB system in the [Fe-S] cluster assembly using oxidative stress and iron depletion as adverse growth conditions. Quantitative real-time polymerase chain reaction demonstrated, for the first time, that Gram-positive bacteria possess an OxyR component responsive to oxidative stress conditions, as fully described for E. coli models. Likewise, strong expression of the sufCDSUB genes was observed in low concentrations of hydrogen peroxide, indicating that the lowest concentration of oxygen free radicals inside cells, known to be highly damaging to [Fe-S] clusters, is sufficient to trigger the transcriptional machinery for prompt replacement of [Fe-S] clusters.
厚壁菌门细菌广泛参与毒力和病理过程。粪肠球菌是一种共生微生物;然而,它也是一种主要与免疫功能低下患者的医院感染相关的病原菌。铁硫[Fe-S]簇是参与多种生物过程的无机辅基,其在体内的形成需要几种特定的蛋白质机制。大肠杆菌是关于[Fe-S]簇生物合成研究最频繁的微生物之一,并编码铁硫簇和硫同化系统。在厚壁菌门物种中,一个由sufCDSUB基因组成的独特操纵子负责[Fe-S]簇的生物合成。本研究的目的是利用氧化应激和铁耗竭作为不利生长条件,研究粪肠球菌sufCDSUB系统在[Fe-S]簇组装中的潜力。定量实时聚合酶链反应首次证明,革兰氏阳性菌拥有一个对氧化应激条件有反应的OxyR成分,这与大肠杆菌模型中所描述的完全一致。同样,在低浓度过氧化氢中观察到sufCDSUB基因的强烈表达,这表明细胞内已知对[Fe-S]簇具有高度破坏性的最低浓度的氧自由基足以触发转录机制,以便迅速替换[Fe-S]簇。