Lee Ai-Yun, Kao Cheng-Yen, Wang Yao-Kuan, Lin Ssu-Yuan, Lai Tze-Ying, Sheu Bor-Shyang, Lo Chien-Jung, Wu Jiunn-Jong
Institute of Molecular Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang Ming University, Taipei, Taiwan.
Helicobacter. 2017 Aug;22(4). doi: 10.1111/hel.12388. Epub 2017 Apr 12.
Flagellar motility of Helicobacter pylori has been shown to be important for the bacteria to establish initial colonization. The ferric uptake regulator (Fur) is a global regulator that has been identified in H. pylori which is involved in the processes of iron uptake and establishing colonization. However, the role of Fur in H. pylori motility is still unclear.
Motility of the wild-type, fur mutant, and fur revertant J99 were determined by a soft-agar motility assay and direct video observation. The bacterial shape and flagellar structure were evaluated by transmission electron microscopy. Single bacterial motility and flagellar switching were observed by phase-contrast microscopy. Autoinducer-2 (AI-2) production in bacterial culture supernatant was analyzed by a bioluminescence assay.
The fur mutant showed impaired motility in the soft-agar assay compared with the wild-type J99 and fur revertant. The numbers and lengths of flagellar filaments on the fur mutant cells were similar to those of the wild-type and revertant cells. Phenotypic characterization showed similar swimming speed but reduction in switching rate in the fur mutant. The AI-2 production of the fur mutant was dramatically reduced compared with wild-type J99 in log-phase culture medium.
These results indicate that Fur positively modulates H. pylori J99 motility through interfering with bacterial flagellar switching.
幽门螺杆菌的鞭毛运动已被证明对该细菌建立初始定植很重要。铁摄取调节因子(Fur)是一种在幽门螺杆菌中已被鉴定出的全局调节因子,它参与铁摄取和建立定植的过程。然而,Fur在幽门螺杆菌运动中的作用仍不清楚。
通过软琼脂运动试验和直接视频观察来测定野生型、fur突变体和fur回复株J99的运动性。通过透射电子显微镜评估细菌形态和鞭毛结构。通过相差显微镜观察单个细菌的运动性和鞭毛切换。通过生物发光试验分析细菌培养上清液中自诱导物-2(AI-2)的产生。
与野生型J99和fur回复株相比,fur突变体在软琼脂试验中的运动性受损。fur突变体细胞上鞭毛丝的数量和长度与野生型和回复株细胞相似。表型特征显示,fur突变体的游动速度相似,但切换率降低。在对数期培养基中,fur突变体的AI-2产量与野生型J99相比显著降低。
这些结果表明,Fur通过干扰细菌鞭毛切换来正向调节幽门螺杆菌J99的运动性。