Kalmokoff Martin, Lanthier Patricia, Tremblay Tammy-Lynn, Foss Mary, Lau Peter C, Sanders Greg, Austin John, Kelly John, Szymanski Christine M
Health Canada Bureau of Microbial Hazards, Ottawa, Ontario K1A 0L2.
J Bacteriol. 2006 Jun;188(12):4312-20. doi: 10.1128/JB.01975-05.
Campylobacter jejuni remains the leading cause of bacterial gastroenteritis in developed countries, and yet little is known concerning the mechanisms by which this fastidious organism survives within its environment. We have demonstrated that C. jejuni 11168 can form biofilms on a variety of surfaces. Proteomic analyses of planktonic and biofilm-grown cells demonstrated differences in protein expression profiles between the two growth modes. Proteins involved in the motility complex, including the flagellins (FlaA, FlaB), the filament cap (FliD), the basal body (FlgG, FlgG2), and the chemotactic protein (CheA), all exhibited higher levels of expression in biofilms than found in stationary-phase planktonic cells. Additional proteins with enhanced expression included those involved in the general (GroEL, GroES) and oxidative (Tpx, Ahp) stress responses, two known adhesins (Peb1, FlaC), and proteins involved in biosynthesis, energy generation, and catabolic functions. An aflagellate flhA mutant not only lost the ability to attach to a solid matrix and form a biofilm but could no longer form a pellicle at the air-liquid interface of a liquid culture. Insertional inactivation of genes that affect the flagellar filament (fliA, flaA, flaB, flaG) or the expression of the cell adhesin (flaC) also resulted in a delay in pellicle formation. These findings demonstrate that the flagellar motility complex plays a crucial role in the initial attachment of C. jejuni 11168 to solid surfaces during biofilm formation as well as in the cell-to-cell interactions required for pellicle formation. Continued expression of the motility complex in mature biofilms is unusual and suggests a role for the flagellar apparatus in the biofilm phenotype.
空肠弯曲菌仍是发达国家细菌性肠胃炎的主要病因，然而对于这种苛求型微生物在其生存环境中的存活机制却知之甚少。我们已经证明，空肠弯曲菌11168能够在多种表面形成生物膜。对浮游生长和生物膜生长细胞的蛋白质组分析表明，两种生长模式下的蛋白质表达谱存在差异。参与运动复合体的蛋白质，包括鞭毛蛋白（FlaA、FlaB）、鞭毛丝帽（FliD）、基体（FlgG、FlgG2）和趋化蛋白（CheA），在生物膜中的表达水平均高于静止期浮游细胞。表达增强的其他蛋白质包括参与一般（GroEL、GroES）和氧化（Tpx、Ahp）应激反应的蛋白质、两种已知的黏附素（Peb1、FlaC）以及参与生物合成、能量产生和分解代谢功能的蛋白质。无鞭毛的flhA突变体不仅失去了附着于固体基质并形成生物膜的能力，而且在液体培养的气液界面也无法再形成菌膜。影响鞭毛丝（fliA、flaA、flaB、flaG）或细胞黏附素（flaC）表达的基因插入失活也导致菌膜形成延迟。这些发现表明，鞭毛运动复合体在空肠弯曲菌11168生物膜形成过程中最初附着于固体表面以及菌膜形成所需的细胞间相互作用中起着至关重要的作用。运动复合体在成熟生物膜中的持续表达并不常见，这表明鞭毛装置在生物膜表型中发挥着作用。
