Expression of the psl operon in Pseudomonas aeruginosa PAO1 biofilms:: PslA performs an essential function in biofilm formation

被引:79
作者
Overhage, J
Schemionek, M
Webb, JS
Rehm, BHA
机构
[1] Massey Univ, Inst Mol Biosci, Palmerston North, New Zealand
[2] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia
[3] Univ New S Wales, Ctr Marine Biofouling & Bioinnovat, Sydney, NSW 2052, Australia
关键词
D O I
10.1128/AEM.71.8.4407-4413.2005
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The psi gene cluster, comprising 15 cotranscribed genes from Pseudomonas aeruginosa, was recently identified as being involved in exopolysaccharide biosynthesis and biofilm formation. In this study, we investigated the regulation of the psi gene cluster and the function of the first gene in this cluster, the pslA gene. PsIA shows strong similarities to UDP-glucose lipid carriers. An isogenic marker-free pslA deletion mutant of P. aeruginosa PAO1 deficient in attachment and biofilm formation was used for complementation studies. The expression (of only the pslA gene, comprising a coding region of 1,437 bp, restored the biofilm-forming phenotype of the wild type, indicating that PslA is required for biofilm formation by nonmucoid P. aeruginosa. The promoter region of the psi gene cluster, which encodes PsIA-PslO, was identified by rapid amplification of cDNA 5' ends. Promoter assays using transcriptional fusions to lacZ and gfp indicated a constitutive expression of the psl cluster in planktonic cells and a highly regulated and localized expression in biofilms, respectively. Expression of the psi cluster in biofilms was almost exclusively found in the centers of microcolonies, as revealed by confocal laser scanning microscopy. These data suggest that constitutive expression of the psl operon enables efficient attachment to surfaces and that regulated localized psi operon expression is required for biofilm differentiation.
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页码:4407 / 4413
页数:7
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