Present address: Imperial College London, Division of Cell and Molecular Biology, Centre for Molecular Microbiology and Infection, South Kensington Campus, Flowers Building, London SW7 2AZ, UK.
The PprA–PprB two-component system activates CupE, the first non-archetypal Pseudomonas aeruginosa chaperone–usher pathway system assembling fimbriae
Version of Record online: 22 NOV 2010
© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
Volume 13, Issue 3, pages 666–683, March 2011
How to Cite
Giraud, C., Bernard, C. S., Calderon, V., Yang, L., Filloux, A., Molin, S., Fichant, G., Bordi, C. and de Bentzmann, S. (2011), The PprA–PprB two-component system activates CupE, the first non-archetypal Pseudomonas aeruginosa chaperone–usher pathway system assembling fimbriae. Environmental Microbiology, 13: 666–683. doi: 10.1111/j.1462-2920.2010.02372.x
- Issue online: 1 MAR 2011
- Version of Record online: 22 NOV 2010
- Received 19 July, 2010; accepted 5 October, 2010.
Fig. S1. Genetic organization of the various cupE gene loci analysed in this study. The following colour scheme was used: pink for cupE1 genes, dark blue for cupE2 genes, orange for cupE3 genes, yellow for cupE4 genes, green for cupE5 genes, light blue for cupE6 genes. The cupE1, cupE2 and cupE3 genes encode fimbrial proteins, cupE4 genes encode chaperone proteins, cupE5 genes encode usherproteins and cupE6 genes encode adhesin-like proteins. The additional copies of a csuA/B gene found only in Pseudomonas species other than P. aeruginosa are coloured in red and the genes encoding spore coat proteins in M. xanthus are shown in purple. Grey boxes indicate genes annotated as pseudogenes in the genome. The asterisk (*) above a white box indicates putative pseudogenes, as the annotation specifies that the frameshift may result from sequence errors. The asterisk (*) above a coloured box indicates a gene that was missed during the annotation but that we identified through a similarity search performed with the tblastn program of the blast suite using, as a query, a protein from another strain of the same species. Species are indicated by the same four-letter code used for phylogenetic trees and the strains are specified: Abau, A. baumannii; Bmal, B. mallei; Bpse, B. pseudomallei; Btha, B. thailandensis; Mxan, M. xanthus; Paer, P. aeruginosa; Pent, P. entomophila; Pflu, P. fluorescens; Pput, P. putida; Spro, S. proteamaculans; Vpar, Vibrio parahaemolyticus; Yent, Y. enterocolitica; Ypes, Y. pestis; Ypse, Y. pseudotuberculosis.
Fig. S2. Phylogenetic trees of the fimbrial proteins (A), the adhesin-like proteins (B) and the 16S rRNA sequences (C), the chaperone proteins (D) and the usher proteins (E). The phylogenetic trees were generated as described in Experimental procedures. The sequences are named according to the nomenclature adopted for P. aeruginosa. We calculated 1000 bootstrap replicates: bootstrap values > 500 are indicated on branches. Trees are coloured on the basis of taxonomy; purple: Enterobacteriales (γ-proteobacteria), green: Pseudomonadaceae (γ-proteobacteria), pink: Burkholderiaceae (β-proteobacteria). Species are indicated by a four-letter code: Abau, A. baumannii; Bmal, B. mallei; Bpse, B. pseudomallei; Btha, B. thailandensis; Mxan, M. xanthus; Paer, P. aeruginosa; Pent: P. entomophila; Pflu, P. fluorescens; Pput, P. putida; Spro, S. proteamaculans; Vpar, V. parahaemolyticus; Yent, Y. enterocolitica, Ypes, Y. pestis.
Fig. S3. Expression of the cupE–lacZ chromosomal fusion was monitored in the PAO1attB::cupE–lacZ parental strain and in clones obtained after transposon mutagenesis in the parental strain on LB and M63 plates. PAO1LBTn20, in which the position of the transposon was mapped upstream from the lacZ gene, with the ptac promoter orientated so as to allow lacZ transcription, was used as a positive control. Expression of cupE–lacZ was also monitored in the PAO1M63Tn17, PAO1M63Tn65, PAO1M63Tn39, PAO1M63Tn25, PAO1M63Tn32, PAO1M63Tn60, PAO1LBTn8, PAO1LBTn11, PAO1LBTn10 and PAO1LBTn24 strains (see Table S1 for transposon localization). Clones corresponding to the same DNA region are indicated with an asterisk.
Table S1. Localization of transposon insertion during mutagenesis of PAO1attB::cupE–lacZ.
Table S2. Strains used in this study.
Table S3. Plasmids used in this study.
Table S4. Oligonucleotides used for mutagenesis and gene cloning.
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