Nitric Oxide Signaling in Pseudomonas aeruginosa Biofilms Mediates Phosphodiesterase Activity, Decreased Cyclic Di-GMP Levels, and Enhanced Dispersal

被引:384
作者
Barraud, Nicolas [1 ,2 ]
Schleheck, David [1 ,2 ]
Klebensberger, Janosch [1 ,2 ]
Webb, Jeremy S. [1 ,2 ]
Hassett, Daniel J. [3 ]
Rice, Scott A. [1 ,2 ]
Kjelleberg, Staffan [1 ,2 ]
机构
[1] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia
[2] Univ New S Wales, Ctr Marine Bioinnovat, Sydney, NSW 2052, Australia
[3] Univ Cincinnati, Coll Med, Dept Mol Genet Biochem & Microbiol, Cincinnati, OH 45267 USA
基金
美国国家卫生研究院; 英国生物技术与生命科学研究理事会; 澳大利亚研究理事会;
关键词
TRANSCRIPTIONAL CONTROL; NITROGEN-OXIDES; EAL DOMAIN; BACTERIAL; EXPRESSION; REGULATOR; PROTEIN; NO; DENITRIFICATION; MOTILITY;
D O I
10.1128/JB.00975-09
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Bacteria in biofilms often undergo active dispersal events and revert to a free-swimming, planktonic state to complete the biofilm life cycle. The signaling molecule nitric oxide (NO) was previously found to trigger biofilm dispersal in the opportunistic pathogen Pseudomonas aeruginosa at low, nontoxic concentrations (N. Barraud, D. J. Hassett, S. H. Hwang, S. A. Rice, S. Kjelleberg, and J. S. Webb, J. Bacteriol. 188: 7344-7353, 2006). NO was further shown to increase cell motility and susceptibility to antimicrobials. Recently, numerous studies revealed that increased degradation of the secondary messenger cyclic di-GMP (c-di-GMP) by specific phosphodiesterases (PDEs) triggers a planktonic mode of growth in eubacteria. In this study, the potential link between NO and c-di-GMP signaling was investigated by performing (i) PDE inhibitor studies, (ii) enzymatic assays to measure PDE activity, and (iii) direct quantification of intracellular c-di-GMP levels. The results suggest a role for c-di-GMP signaling in triggering the biofilm dispersal event induced by NO, as dispersal requires PDE activity and addition of NO stimulates PDE and induces the concomitant decrease in intracellular c-di-GMP levels in P. aeruginosa. Furthermore, gene expression studies indicated global responses to low, nontoxic levels of NO in P. aeruginosa biofilms, including upregulation of genes involved in motility and energy metabolism and downregulation of adhesins and virulence factors. Finally, site-directed mutagenesis of candidate genes and physiological characterization of the corresponding mutant strains uncovered that the chemotaxis transducer BdlA is involved in the biofilm dispersal response induced by NO.
引用
收藏
页码:7333 / 7342
页数:10
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