Swarming motility is modulated by expression of the putative xenosiderophore transporter SppR-SppABCD in Pseudomonas aeruginosa PA14
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In the present study, we characterised the putative peptide ABC transporter SppABCD, which is co-transcribed with the TonB-dependent receptor SppR in Pseudomonas aeruginosa PA14. However, our data show that this transporter complex is not involved in the uptake of peptides. The fact that the TonB-dependent receptor SppR is regulated by an iron starvation ECF sigma factor suggested that this transporter is probably involved in the uptake of xenosiderophores. Therefore, we screened culture supernatants of 23 siderophore-producing bacteria for their ability to induce the expression of the SppR-regulating ECF sigma factor. However, none of them had an effect on the expression of this ECF sigma factor. Since the spp operon is not expressed under standard laboratory conditions, we overexpressed it from plasmids in PA14, which led to an impairment of its swarming motility on semisolid agar. Since we excluded the possibility that the uptake of a culture medium component was responsible for the observed phenotype, we hypothesize that the Spp transport system is involved in the uptake of a compound from the periplasmic space or a compound secreted by P. aeruginosa. Furthermore, we found that rhamnolipid synthesis was decreased while biofilm and exopolysaccharide synthesis was slightly increased upon overexpression of the spp operon. Moreover, we observed an impact of spp overexpression on regulation of genes involved in siderophore and phenazine biosynthesis.
KeywordsBiofilm Exopolysaccharide Pyoverdine Phenazine Rhamnolipid Siderophore
We gratefully acknowledge Svetlana Dubiley and Konstantin Severinov for experimental testing of Microcin C uptake. Thanks to Daniel Boland for pre-screening environmental siderophore-producing bacteria. We appreciate the help of Michael Mourez for Biolog experiments. Furthermore, we acknowledge Mathias Winterhalter, Roland Benz, Thilo Köhler, and Malcolm Page for generous laboratory support and useful discussions.
The research leading to these results has received support from the Innovative Medicines Initiatives Joint Undertaking under Grant Agreement No. 115525, resources which are composed of financial contributions from the European Union’s seventh framework programme (FP7/2007-2013) and European Federation of Pharmaceutical Industries and Associations companies in kind contribution.
- Choi KH, Kumar A, Schweizer HP (2006) A 10-min method for preparation of highly electrocompetent Pseudomonas aeruginosa cells: application for DNA fragment transfer between chromosomes and plasmid transformation. J Microbiol Methods 64:391–397. doi: 10.1016/j.mimet.2005.06.001 CrossRefPubMedGoogle Scholar
- Clinical and Laboratory Standards Institute (2012) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. In. CLSI document M7-A7, Clinical and Laboratory Standards Institute, Wayne, PAGoogle Scholar
- Mahren S, Enz S, Braun V (2002) Functional interaction of region 4 of the extracytoplasmic function sigma factor FecI with the cytoplasmic portion of the FecR transmembrane protein of the Escherichia coli ferric citrate transport system. J Bacteriol 184:3704–3711CrossRefPubMedPubMedCentralGoogle Scholar
- Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
- Murray TS, Ledizet M, Kazmierczak BI (2010) Swarming motility, secretion of type 3 effectors and biofilm formation phenotypes exhibited within a large cohort of Pseudomonas aeruginosa clinical isolates. J Med Microbiol 59:511–520. doi: 10.1099/jmm.0.017715-0 CrossRefPubMedPubMedCentralGoogle Scholar
- Schenk A, Weingart H, Ullrich MS (2008) Extraction of high-quality bacterial RNA from infected leaf tissue for bacterial in planta gene expression analysis by multiplexed fluorescent Northern hybridization. Mol Plant Pathol 9:227–235. doi: 10.1111/j.1364-3703.2007.00452.x CrossRefPubMedGoogle Scholar
- Schweizer HP, Chuanchuen R (2001) Small broad-host-range lacZ operon fusion vector with low background activity. Biotechniques 31:1258, 1260, 1262Google Scholar