van Schaik Erin J, Giltner Carmen L, Audette Gerald F, Keizer David W, Bautista Daisy L, Slupsky Carolyn M, Sykes Brian D, Irvin Randall T
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
J Bacteriol. 2005 Feb;187(4):1455-64. doi: 10.1128/JB.187.4.1455-1464.2005.
The opportunistic pathogen Pseudomonas aeruginosa produces multifunctional, polar, filamentous appendages termed type IV pili. Type IV pili are involved in colonization during infection, twitching motility, biofilm formation, bacteriophage infection, and natural transformation. Electrostatic surface analysis of modeled pilus fibers generated from P. aeruginosa strain PAK, K122-4, and KB-7 pilin monomers suggested that a solvent-exposed band of positive charge may be a common feature of all type IV pili. Several functions of type IV pili, including natural transformation and biofilm formation, involve DNA. We investigated the ability of P. aeruginosa type IV pili to bind DNA. Purified PAK, K122-4, and KB-7 pili were observed to bind both bacterial plasmid and salmon sperm DNA in a concentration-dependent and saturable manner. PAK pili had the highest affinity for DNA, followed by K122-4 and KB-7 pili. DNA binding involved backbone interactions and preferential binding to pyrimidine residues even though there was no evidence of sequence-specific binding. Pilus-mediated DNA binding was a function of the intact pilus and thus required elements present in the quaternary structure. However, binding also involved the pilus tip as tip-specific, but not base-specific, antibodies inhibited DNA binding. The conservation of a Thr residue in all type IV pilin monomers examined to date, along with the electrostatic data, implies that DNA binding is a conserved function of type IV pili. Pilus-mediated DNA binding could be important for biofilm formation both in vivo during an infection and ex vivo on abiotic surfaces.
机会致病菌铜绿假单胞菌产生称为IV型菌毛的多功能、极性、丝状附属物。IV型菌毛参与感染期间的定殖、颤动运动、生物膜形成、噬菌体感染和自然转化。对由铜绿假单胞菌PAK菌株、K122 - 4和KB - 7菌毛单体生成的模拟菌毛纤维进行的静电表面分析表明,溶剂暴露的正电荷带可能是所有IV型菌毛的共同特征。IV型菌毛的几种功能,包括自然转化和生物膜形成,都涉及DNA。我们研究了铜绿假单胞菌IV型菌毛结合DNA的能力。观察到纯化的PAK、K122 - 4和KB - 7菌毛以浓度依赖性和饱和方式结合细菌质粒和鲑鱼精DNA。PAK菌毛对DNA的亲和力最高,其次是K122 - 4和KB - 7菌毛。DNA结合涉及主链相互作用以及对嘧啶残基的优先结合,尽管没有序列特异性结合的证据。菌毛介导的DNA结合是完整菌毛的功能,因此需要四级结构中存在的元件。然而,结合也涉及菌毛尖端,因为尖端特异性而非基部特异性抗体抑制DNA结合。迄今为止在所有检测的IV型菌毛单体中Thr残基的保守性,连同静电数据,意味着DNA结合是IV型菌毛的保守功能。菌毛介导的DNA结合对于感染期间体内和非生物表面体外的生物膜形成可能都很重要。