College of Life Sciences, Nankai University, Tianjin, 300071, China; School of Public Health, Chongqing Medical University, Chongqing, 400016, China.
College of Life Sciences, Nankai University, Tianjin, 300071, China.
Microb Pathog. 2020 Jul;144:103982. doi: 10.1016/j.micpath.2020.103982. Epub 2020 Feb 24.
Pseudomonas aeruginosa is a nosocomial human pathogen causing infections in immunocompromised patients. To explore new genes involved in P. aeruginosa swimming motility, Mu transposon mutagenesis library was screened for isolates with altered swimming motility. Eleven nonmobile mutants were identified. Sequence analysis shows the nonmobile phenotype of one isolate was attributed to the inactivation of PA5001 gene. PA5001 knockout mutant based on the PAK lab strain also displayed comparable phenotypes suggesting the universal gene function regardless of strain. Exotic PA5001 gene fragment provided on expressing plasmid was capable of storing nonmobile phenotype of PA5001 mutant, suggesting the functional involvement of PA5001 gene on bacterial swimming. Impact of PA5001 inactivation on biofilm formation was examined, as adhesion and interaction during biofilm formation is highly dependent of bacterial mobility. The result shows that normal architecture of biofilm was disrupted in the mutant. Complementing by exotic PA5001 gene fragment resulted in the restoration of biofilm phenotype. Our results provide evidences suggesting the functional participation of PA5001 gene in bacterial mobility and biofilm formation. The critical function by PA5001 in bacterial motility and biofilm might serve as hint for the novel target for the treatment of chronic infections caused by P. aeruginosa.
铜绿假单胞菌是一种医院获得性人类病原体,可导致免疫功能低下患者感染。为了探索参与铜绿假单胞菌游动性的新基因,我们筛选了游动性发生改变的 Mu 转座子突变体文库。鉴定出 11 个非运动突变体。序列分析表明,一个分离株的非运动表型归因于 PA5001 基因的失活。基于 PAK 实验室菌株的 PA5001 敲除突变体也显示出类似的表型,这表明该基因的普遍功能与菌株无关。表达质粒上提供的外来 PA5001 基因片段能够储存 PA5001 突变体的非运动表型,这表明 PA5001 基因在细菌游动中具有功能作用。我们还研究了 PA5001 失活对生物膜形成的影响,因为生物膜形成过程中的粘附和相互作用高度依赖于细菌的流动性。结果表明,突变体中生物膜的正常结构被破坏。通过外来 PA5001 基因片段的互补,恢复了生物膜表型。我们的结果提供了证据,表明 PA5001 基因在细菌运动性和生物膜形成中具有功能参与。PA5001 在细菌运动和生物膜中的关键功能可能为治疗铜绿假单胞菌引起的慢性感染提供新的治疗靶点。