Pseudomonas aeruginosa uses a cyclic-di-GMP-regulated adhesin to reinforce the biofilm extracellular matrix

被引:411
作者
Borlee, Bradley R. [1 ]
Goldman, Aaron D. [1 ]
Murakami, Keiji [1 ]
Samudrala, Ram [1 ]
Wozniak, Daniel J. [2 ]
Parsek, Matthew R. [1 ]
机构
[1] Univ Washington, Dept Microbiol, Seattle, WA 98195 USA
[2] Ohio State Univ, Ctr Microbial Interface Biol, Columbus, OH 43210 USA
基金
美国国家科学基金会;
关键词
FILAMENTOUS HEMAGGLUTININ ADHESIN; OUTER-MEMBRANE PROTEIN; SMALL-COLONY VARIANTS; ESCHERICHIA-COLI; BORDETELLA-PERTUSSIS; ANTIGEN; 43; VIRULENCE FACTORS; GENES; PSL; EXOPOLYSACCHARIDE;
D O I
10.1111/j.1365-2958.2009.06991.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
P>Pseudomonas aeruginosa, the principal pathogen of cystic fibrosis patients, forms antibiotic-resistant biofilms promoting chronic colonization of the airways. The extracellular (EPS) matrix is a crucial component of biofilms that provides the community multiple benefits. Recent work suggests that the secondary messenger, cyclic-di-GMP, promotes biofilm formation. An analysis of factors specifically expressed in P. aeruginosa under conditions of elevated c-di-GMP, revealed functions involved in the production and maintenance of the biofilm extracellular matrix. We have characterized one of these components, encoded by the PA4625 gene, as a putative adhesin and designated it cdrA. CdrA shares structural similarities to extracellular adhesins that belong to two-partner secretion systems. The cdrA gene is in a two gene operon that also encodes a putative outer membrane transporter, CdrB. The cdrA gene encodes a 220 KDa protein that is predicted to be rod-shaped protein harbouring a beta-helix structural motif. Western analysis indicates that the CdrA is produced as a 220 kDa proprotein and processed to 150 kDa before secretion into the extracellular medium. We demonstrated that cdrAB expression is minimal in liquid culture, but is elevated in biofilm cultures. CdrAB expression was found to promote biofilm formation and auto-aggregation in liquid culture. Aggregation mediated by CdrA is dependent on the Psl polysaccharide and can be disrupted by adding mannose, a key structural component of Psl. Immunoprecipitation of Psl present in culture supernatants resulted in co-immunoprecipitation of CdrA, providing additional evidence that CdrA directly binds to Psl. A mutation in cdrA caused a decrease in biofilm biomass and resulted in the formation of biofilms exhibiting decreased structural integrity. Psl-specific lectin staining suggests that CdrA either cross-links Psl polysaccharide polymers and/or tethers Psl to the cells, resulting in increased biofilm structural stability. Thus, this study identifies a key protein structural component of the P. aeruginosa EPS matrix.
引用
收藏
页码:827 / 842
页数:16
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