Liu Jia, Guo Lihong, Liu Jianwei, Zhang Jianying, Zeng Huihui, Ning Yang, Wei Xi
Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen UniversityGuangzhou, China.
Department of Operative Dentistry and Endodontics, Xiangya Stomatological Hospital, Central South UniversityChangsha, China.
Front Microbiol. 2017 Jun 8;8:962. doi: 10.3389/fmicb.2017.00962. eCollection 2017.
Efflux transporters have been implicated in regulating bacterial virulence properties such as resistance to antibiotics, biofilm formation and colonization. The pathogenicity of , the primary etiologic agent of human dental caries, relies on the bacterium's ability to form biofilms on tooth surface. However, the studies on efflux transporters in are scare and the function of these transporters remained to be clarified. In this study, we identified an efflux transporter (LmrB) in through cloning the gene into . Introducing into conferred a multidrug-resistant phenotype and resulted in higher EtBr efflux activity which could be suppressed by efflux inhibitor. To explore whether LmrB was involved in virulence properties regulation, we constructed the inactivation mutant and examined the phenotypes of the mutant. It was found that LmrB deficiency resulted in increased IPS storage and prolonged acid production. Enhanced biofilm formation characterized by increased extracellular polysaccharides (EPS) production and elevated resistance to hydrogen peroxide and antimicrobials were also observed in mutant. To gain a better understanding of the global role of LmrB, a transcriptome analysis was performed using mutant strain. The expression of 107 genes was up- or down-regulated in the mutant compared with the wild type. Notably, expression of genes in several genomic islands was differentially modulated, such as stress-related GroELS and scnRK, sugar metabolism associated operons and transporter. The results presented here indicate that LmrB plays a vital global role in the regulation of several important virulence properties in .
外排转运蛋白与调节细菌毒力特性有关,如抗生素耐药性、生物膜形成和定植。变形链球菌是人类龋齿的主要病原体,其致病性依赖于该细菌在牙齿表面形成生物膜的能力。然而,关于变形链球菌中外排转运蛋白的研究很少,这些转运蛋白的功能仍有待阐明。在本研究中,我们通过将变形链球菌的lmrB基因克隆到大肠杆菌中,鉴定出一种外排转运蛋白(LmrB)。将LmrB导入大肠杆菌赋予了多药耐药表型,并导致更高的溴化乙锭外排活性,该活性可被外排抑制剂抑制。为了探究LmrB是否参与变形链球菌毒力特性的调节,我们构建了lmrB失活突变体并检测了该突变体的表型。结果发现,LmrB缺陷导致细胞内多糖储存增加和产酸时间延长。在lmrB突变体中还观察到生物膜形成增强,其特征是细胞外多糖(EPS)产量增加,以及对过氧化氢和抗菌剂的耐受性增强。为了更好地理解LmrB的全局作用,我们使用lmrB突变株进行了转录组分析。与野生型相比,lmrB突变体中有107个基因的表达上调或下调。值得注意的是,几个基因组岛中的基因表达受到差异调节,如与应激相关的GroELS和scnRK、与糖代谢相关的操纵子和转运蛋白。此处呈现的结果表明,LmrB在调节变形链球菌的几种重要毒力特性中起着至关重要的全局作用。