Blanco P, Corona F, Sánchez M B, Martínez J L
Centro Nacional de Biotecnología, CSIC, Madrid, Spain.
Centro Nacional de Biotecnología, CSIC, Madrid, Spain
Antimicrob Agents Chemother. 2017 Apr 24;61(5). doi: 10.1128/AAC.02453-16. Print 2017 May.
is an opportunistic pathogen with increasing prevalence, which is able to cause infections in immunocompromised patients or in those with a previous pathology. The treatment of the infections caused by this bacterium is often complicated due to the several intrinsic antibiotic resistance mechanisms that it presents. Multidrug efflux pumps are among the best-studied mechanisms of antibiotic resistance. Some of these efflux pumps have a basal expression level but, in general, their expression is often low and only reaches high levels when the local regulator is mutated or bacteria are in the presence of an effector. In the current work, we have developed a yellow fluorescent protein (YFP)-based sensor with the aim to identify effectors able to trigger the expression of SmeVWX, an efflux pump that confers resistance to quinolones, chloramphenicol, and tetracycline when it is expressed at high levels. With this purpose in mind, we tested a variety of different compounds and analyzed the fluorescence signal given by the expression of YFP under the control of the promoter. Among the tested compounds, vitamin K, which is a compound belonging to the 2-methyl-1,4-naphthoquinone family, is produced by plants in defense against infection, and has increasing importance in human therapy, was able to induce the expression of the SmeVWX efflux pump. In addition, a decrease in the susceptibility of to ofloxacin and chloramphenicol was observed in the presence of vitamin K, in both wild-type and -deficient strains.
是一种患病率不断上升的机会致病菌,能够在免疫功能低下的患者或有既往病史的患者中引起感染。由于该细菌呈现出多种内在的抗生素耐药机制,其所致感染的治疗往往很复杂。多药外排泵是研究得最为深入的抗生素耐药机制之一。其中一些外排泵有基础表达水平,但一般来说,它们的表达通常较低,只有当局部调节因子发生突变或细菌处于效应物存在的情况下才会达到高水平。在当前的工作中,我们开发了一种基于黄色荧光蛋白(YFP)的传感器,旨在识别能够触发SmeVWX表达的效应物,SmeVWX是一种外排泵,当它高水平表达时会赋予对喹诺酮类、氯霉素和四环素的耐药性。出于这个目的,我们测试了各种不同的化合物,并分析了在启动子控制下YFP表达所给出的荧光信号。在测试的化合物中,维生素K是一种属于2-甲基-1,4-萘醌家族的化合物,由植物产生用于抵御感染,并且在人类治疗中的重要性日益增加,它能够诱导SmeVWX外排泵的表达。此外,在野生型和缺陷型菌株中,在维生素K存在的情况下,观察到对氧氟沙星和氯霉素的敏感性降低。