Lo Yi-Ling, Shen Lunda, Chang Chih-Hsuan, Bhuwan Manish, Chiu Cheng-Hsun, Chang Hwan-You
Institute of Molecular Medicine, National Tsing Hua University, Hsin Chu, Taiwan.
Molecular Infectious Disease Research Center, Division of Pediatric Infectious Diseases, Department of Pediatrics, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taoyuan, Taiwan.
PLoS One. 2016 May 13;11(5):e0155397. doi: 10.1371/journal.pone.0155397. eCollection 2016.
The transcription factor FliA, also called sigma 28, is a major regulator of bacterial flagellar biosynthesis genes. Growing evidence suggest that in addition to motility, FliA is involved in controlling numerous bacterial behaviors, even though the underlying regulatory mechanism remains unclear. By using a transcriptional fusion to gfp that responds to cyclic (c)-di-GMP, this study revealed a higher c-di-GMP concentration in the fliA deletion mutant of Pseudomonas aeruginosa than in its wild-type strain PAO1. A comparative analysis of transcriptome profiles of P. aeruginosa PAO1 and its fliA deletion mutant revealed an altered expression of several c-di-GMP-modulating enzyme-encoding genes in the fliA deletion mutant. Moreover, the downregulation of PA4367 (bifA), a Glu-Ala-Leu motif-containing phosphodiesterase, in the fliA deletion mutant was confirmed using the β-glucuronidase reporter gene assay. FliA also altered pyocyanin and pyorubin production by modulating the c-di-GMP concentration. Complementing the fliA mutant strain with bifA restored the motility defect and pigment overproduction of the fliA mutant. Our results indicate that in addition to regulating flagellar gene transcription, FliA can modulate the c-di-GMP concentration to regulate the swarming motility and phenazine pigment production in P. aeruginosa.
转录因子FliA,也称为σ28,是细菌鞭毛生物合成基因的主要调节因子。越来越多的证据表明,除了运动性之外,FliA还参与控制多种细菌行为,尽管其潜在的调控机制尚不清楚。通过使用对环(c)-二鸟苷单磷酸(c-di-GMP)有反应的gfp转录融合,本研究发现铜绿假单胞菌fliA缺失突变体中的c-di-GMP浓度高于其野生型菌株PAO1。对铜绿假单胞菌PAO1及其fliA缺失突变体的转录组图谱进行比较分析,发现fliA缺失突变体中几个编码c-di-GMP调节酶的基因表达发生了改变。此外,使用β-葡萄糖醛酸酶报告基因检测证实了fliA缺失突变体中PA4367(bifA)的下调,PA4367是一种含有Glu-Ala-Leu基序的磷酸二酯酶。FliA还通过调节c-di-GMP浓度改变了绿脓菌素和脓青素的产生。用bifA补充fliA突变菌株可恢复fliA突变体的运动缺陷和色素过度产生。我们的结果表明,除了调节鞭毛基因转录外,FliA还可以调节c-di-GMP浓度,以调节铜绿假单胞菌的群体运动和吩嗪色素产生。