Jenkins A Toby A, Buckling Angus, McGhee Marsha, ffrench-Constant Richard H
Departments of Chemistry and Biology, University of Bath, Bath BA2 7AY, UK.
J R Soc Interface. 2005 Jun 22;2(3):255-9. doi: 10.1098/rsif.2005.0030.
Type IV pili have been shown to play a role in the early stages of bacterial biofilm formation, but not in initial bacterial attachment. Here, using the surface analytical technique, surface plasmon resonance (SPR), we follow the attachment of the bacterium Pseudomonas aeruginosa in real time. In contrast to previous studies, we show that type IV pili mutants are defective in attachment. Both mutants lacking pili (pilA), and those possessing an overabundance of pili (pilT), showed reduced SPR measured attachment compared with the wild-type PAO1 strain. Both pil mutants also showed reduced pathogenicity in a model insect host, as measured by percentage mortality after 24h. SPR revealed differences in the kinetics of attachment between pilA and pilT, differences obscured by endpoint assays using crystal violet stain. These results highlight the power of SPR in monitoring bacterial attachment in real time and also demonstrate an additional role for type IV pili beyond bacterial aggregation and micro-colony formation.
IV型菌毛已被证明在细菌生物膜形成的早期阶段发挥作用,但在细菌初始附着过程中并非如此。在此,我们使用表面分析技术表面等离子体共振(SPR)实时跟踪铜绿假单胞菌的附着情况。与之前的研究不同,我们发现IV型菌毛突变体在附着方面存在缺陷。与野生型PAO1菌株相比,缺乏菌毛的突变体(pilA)和菌毛过多的突变体(pilT)的SPR测量附着率均降低。通过24小时后的死亡率百分比测量,这两种pil突变体在模型昆虫宿主中的致病性也均降低。SPR揭示了pilA和pilT在附着动力学上的差异,而使用结晶紫染色的终点测定法掩盖了这些差异。这些结果突出了SPR在实时监测细菌附着方面的强大功能,也证明了IV型菌毛在细菌聚集和微菌落形成之外的额外作用。