Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
J Bacteriol. 2011 Jun;193(11):2717-25. doi: 10.1128/JB.00149-11. Epub 2011 Mar 25.
Organic compounds exhibit various levels of toxicity toward living organisms based upon their ability to insert into biological membranes and disrupt normal membrane function. The primary mechanism responsible for organic solvent tolerance in many bacteria is energy-dependent extrusion via efflux pumps. One such bacterial strain, Pseudomonas putida S12, is known for its high tolerance to organic solvents as provided through the SrpABC resistance-nodulation-cell division (RND) family efflux pump. To determine how two putative regulatory proteins (SrpR and SrpS, encoded directly upstream of the SrpABC structural genes) influence SrpABC efflux pump expression, we conducted transcriptional analysis, β-galactosidase fusion experiments, electrophoretic mobility shift assays, and pulldown analysis. Together, the results of these experiments suggest that expression of the srpABC operon can be derepressed by two distinct but complementary mechanisms: direct inhibition of the SrpS repressor by organic solvents and binding of SrpS by its antirepressor SrpR.
有机化合物根据其插入生物膜并破坏正常膜功能的能力,对生物体表现出不同程度的毒性。许多细菌对有机溶剂的耐受性的主要机制是通过外排泵进行能量依赖性的排出。一种这样的细菌菌株,恶臭假单胞菌 S12,以其通过 SrpABC 抗性-结节-细胞分裂(RND)家族外排泵提供的高有机溶剂耐受性而闻名。为了确定两个假定的调节蛋白(SrpR 和 SrpS,直接在 SrpABC 结构基因的上游编码)如何影响 SrpABC 外排泵的表达,我们进行了转录分析、β-半乳糖苷酶融合实验、电泳迁移率变动分析和下拉分析。这些实验的结果表明,srpABC 操纵子的表达可以通过两种不同但互补的机制被解除抑制:有机溶剂直接抑制 SrpS 阻遏物和 SrpR 结合其抗阻遏物 SrpS。