Limits...
O serotype-independent susceptibility of Pseudomonas aeruginosa to lectin-like pyocins.

Ghequire MG, Dingemans J, Pirnay JP, De Vos D, Cornelis P, De Mot R - Microbiologyopen (2014)

Bottom Line: The recombinant proteins exhibit species-specific antagonistic activities down to nanomolar concentrations against clinical and environmental P. aeruginosa strains, including several multidrug-resistant isolates.No correlation was found between L pyocin susceptibility and phylogenetic relatedness of P. aeruginosa isolates.Sensitive strains were retrieved in 13 out of 15 O serotypes tested, excluding the possibility that the highly variable and immunogenic O serotype antigen of the LPS coating would represent a dominant susceptibility-discriminating factor.

View Article: PubMed Central - PubMed

Affiliation: Centre of Microbial and Plant Genetics, University of Leuven, 3001, Heverlee, Belgium.

Show MeSH

Related in: MedlinePlus

Phylogeny of Pseudomonas aeruginosa proteins with tandem-monocot mannose-binding lectin (MMBL) architecture. (A) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment (Fig. S1) of bacteriocins from P. aeruginosa C1433, Pseudomonas putida BW11M1, Pseudomonas protegens Pf-5 (a, PFL_1229; b, PFL_2127), Pseudomonas syringae pv. syringae 642, X. axonopodis pv. citri str. 306, B. cenocepacia AU1054, with hypothetical proteins from P. aeruginosa BWHPSA007 (ERZ04935), P. aeruginosa 62 (ERX71449), P. chlororaphis 30-84 (EJL08681), Pseudomonas fluorescens NCIMB 11764 (ERP40244), P. syringae DSM50252 (EGH77666), Pseudomonas sp. GM80 (a, EJN36166; b, EJN36323) and R62 (derived from AHZM01000533). Orthologues sharing >95% amino acid identity with one of the listed proteins are not included in the tree and Figure S1. Predicted amino-terminal signal peptide sequences were removed before alignment (names marked with asterisk). Pairs of tandem MMBL proteins present in a particular strain are indicated with extensions (a) and (b). The scale bar represents 0.4 substitutions per site. The positions of the C1433 and BWHPSA007 pyocin groups are highlighted in green and purple, respectively. (B) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment of individual MMBL modules extracted from Pseudomonas tandem-MMBL proteins (Fig. S1). Protein sequences and designations are the same as used in (A). The P. aeruginosa amino-terminal (N, red) and carboxy-terminal-domains (C, blue) are highlighted. The scale bar represents 0.5 substitutions per site. Bootstrap values (percentage of 100 replicates) higher than or equal to 50 are shown at the branches.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4263511&req=5

fig01: Phylogeny of Pseudomonas aeruginosa proteins with tandem-monocot mannose-binding lectin (MMBL) architecture. (A) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment (Fig. S1) of bacteriocins from P. aeruginosa C1433, Pseudomonas putida BW11M1, Pseudomonas protegens Pf-5 (a, PFL_1229; b, PFL_2127), Pseudomonas syringae pv. syringae 642, X. axonopodis pv. citri str. 306, B. cenocepacia AU1054, with hypothetical proteins from P. aeruginosa BWHPSA007 (ERZ04935), P. aeruginosa 62 (ERX71449), P. chlororaphis 30-84 (EJL08681), Pseudomonas fluorescens NCIMB 11764 (ERP40244), P. syringae DSM50252 (EGH77666), Pseudomonas sp. GM80 (a, EJN36166; b, EJN36323) and R62 (derived from AHZM01000533). Orthologues sharing >95% amino acid identity with one of the listed proteins are not included in the tree and Figure S1. Predicted amino-terminal signal peptide sequences were removed before alignment (names marked with asterisk). Pairs of tandem MMBL proteins present in a particular strain are indicated with extensions (a) and (b). The scale bar represents 0.4 substitutions per site. The positions of the C1433 and BWHPSA007 pyocin groups are highlighted in green and purple, respectively. (B) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment of individual MMBL modules extracted from Pseudomonas tandem-MMBL proteins (Fig. S1). Protein sequences and designations are the same as used in (A). The P. aeruginosa amino-terminal (N, red) and carboxy-terminal-domains (C, blue) are highlighted. The scale bar represents 0.5 substitutions per site. Bootstrap values (percentage of 100 replicates) higher than or equal to 50 are shown at the branches.

Mentions: Multiple genes encoding proteins with a tandem-MMBL domain architecture were retrieved in several P. aeruginosa genomic sequences. Being only distantly related to LlpAs of other Pseudomonas species (<30% amino acid sequence identity), these proteins fall into two remote groups with borderline pairwise homology (˜28% identity) (Fig. 1A, Fig. S1, Table S3). One cluster consists of PyoL1 homologues with sequences either identical to the C1433 protein (in strain C1426, the non-mucoid progenitor of mucoid strain C1433) (Stewart et al. 2014) or sharing >86% identity to it (in strains 62, 3578, 3579, BWHPSA001, and BWHPSA016; Pseudomonas aeruginosa initiative, Broad Institute). Within the newly identified second cluster, pairwise sequence identities between the proteins from strains BK1 (Jeganathan et al. 2014), BWHPSA007 (with two homologues) and BWH033 (Pseudomonas aeruginosa initiative, Broad Institute), and VRFPA06 (Murugan et al. 2014) exceed 95%. The members of the respective clusters also differ by the presence (BWHPSA007 group) or absence (C1433 group) of an amino-terminal Sec-specific signal sequence. In this respect, PyoL1 and its homologues resemble other Pseudomonas LlpAs, lacking a discernable secretion motif, whereas the members of the second group are most likely substrates of the Sec translocon, similarly to Xanthomonas and Burkholderia LlpAs (Ghequire et al. 2012b, 2013b).


O serotype-independent susceptibility of Pseudomonas aeruginosa to lectin-like pyocins.

Ghequire MG, Dingemans J, Pirnay JP, De Vos D, Cornelis P, De Mot R - Microbiologyopen (2014)

Phylogeny of Pseudomonas aeruginosa proteins with tandem-monocot mannose-binding lectin (MMBL) architecture. (A) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment (Fig. S1) of bacteriocins from P. aeruginosa C1433, Pseudomonas putida BW11M1, Pseudomonas protegens Pf-5 (a, PFL_1229; b, PFL_2127), Pseudomonas syringae pv. syringae 642, X. axonopodis pv. citri str. 306, B. cenocepacia AU1054, with hypothetical proteins from P. aeruginosa BWHPSA007 (ERZ04935), P. aeruginosa 62 (ERX71449), P. chlororaphis 30-84 (EJL08681), Pseudomonas fluorescens NCIMB 11764 (ERP40244), P. syringae DSM50252 (EGH77666), Pseudomonas sp. GM80 (a, EJN36166; b, EJN36323) and R62 (derived from AHZM01000533). Orthologues sharing >95% amino acid identity with one of the listed proteins are not included in the tree and Figure S1. Predicted amino-terminal signal peptide sequences were removed before alignment (names marked with asterisk). Pairs of tandem MMBL proteins present in a particular strain are indicated with extensions (a) and (b). The scale bar represents 0.4 substitutions per site. The positions of the C1433 and BWHPSA007 pyocin groups are highlighted in green and purple, respectively. (B) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment of individual MMBL modules extracted from Pseudomonas tandem-MMBL proteins (Fig. S1). Protein sequences and designations are the same as used in (A). The P. aeruginosa amino-terminal (N, red) and carboxy-terminal-domains (C, blue) are highlighted. The scale bar represents 0.5 substitutions per site. Bootstrap values (percentage of 100 replicates) higher than or equal to 50 are shown at the branches.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263511&req=5

fig01: Phylogeny of Pseudomonas aeruginosa proteins with tandem-monocot mannose-binding lectin (MMBL) architecture. (A) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment (Fig. S1) of bacteriocins from P. aeruginosa C1433, Pseudomonas putida BW11M1, Pseudomonas protegens Pf-5 (a, PFL_1229; b, PFL_2127), Pseudomonas syringae pv. syringae 642, X. axonopodis pv. citri str. 306, B. cenocepacia AU1054, with hypothetical proteins from P. aeruginosa BWHPSA007 (ERZ04935), P. aeruginosa 62 (ERX71449), P. chlororaphis 30-84 (EJL08681), Pseudomonas fluorescens NCIMB 11764 (ERP40244), P. syringae DSM50252 (EGH77666), Pseudomonas sp. GM80 (a, EJN36166; b, EJN36323) and R62 (derived from AHZM01000533). Orthologues sharing >95% amino acid identity with one of the listed proteins are not included in the tree and Figure S1. Predicted amino-terminal signal peptide sequences were removed before alignment (names marked with asterisk). Pairs of tandem MMBL proteins present in a particular strain are indicated with extensions (a) and (b). The scale bar represents 0.4 substitutions per site. The positions of the C1433 and BWHPSA007 pyocin groups are highlighted in green and purple, respectively. (B) Unrooted maximum-likelihood tree inferred from a multiple amino acid sequence alignment of individual MMBL modules extracted from Pseudomonas tandem-MMBL proteins (Fig. S1). Protein sequences and designations are the same as used in (A). The P. aeruginosa amino-terminal (N, red) and carboxy-terminal-domains (C, blue) are highlighted. The scale bar represents 0.5 substitutions per site. Bootstrap values (percentage of 100 replicates) higher than or equal to 50 are shown at the branches.
Mentions: Multiple genes encoding proteins with a tandem-MMBL domain architecture were retrieved in several P. aeruginosa genomic sequences. Being only distantly related to LlpAs of other Pseudomonas species (<30% amino acid sequence identity), these proteins fall into two remote groups with borderline pairwise homology (˜28% identity) (Fig. 1A, Fig. S1, Table S3). One cluster consists of PyoL1 homologues with sequences either identical to the C1433 protein (in strain C1426, the non-mucoid progenitor of mucoid strain C1433) (Stewart et al. 2014) or sharing >86% identity to it (in strains 62, 3578, 3579, BWHPSA001, and BWHPSA016; Pseudomonas aeruginosa initiative, Broad Institute). Within the newly identified second cluster, pairwise sequence identities between the proteins from strains BK1 (Jeganathan et al. 2014), BWHPSA007 (with two homologues) and BWH033 (Pseudomonas aeruginosa initiative, Broad Institute), and VRFPA06 (Murugan et al. 2014) exceed 95%. The members of the respective clusters also differ by the presence (BWHPSA007 group) or absence (C1433 group) of an amino-terminal Sec-specific signal sequence. In this respect, PyoL1 and its homologues resemble other Pseudomonas LlpAs, lacking a discernable secretion motif, whereas the members of the second group are most likely substrates of the Sec translocon, similarly to Xanthomonas and Burkholderia LlpAs (Ghequire et al. 2012b, 2013b).

Bottom Line: The recombinant proteins exhibit species-specific antagonistic activities down to nanomolar concentrations against clinical and environmental P. aeruginosa strains, including several multidrug-resistant isolates.No correlation was found between L pyocin susceptibility and phylogenetic relatedness of P. aeruginosa isolates.Sensitive strains were retrieved in 13 out of 15 O serotypes tested, excluding the possibility that the highly variable and immunogenic O serotype antigen of the LPS coating would represent a dominant susceptibility-discriminating factor.

View Article: PubMed Central - PubMed

Affiliation: Centre of Microbial and Plant Genetics, University of Leuven, 3001, Heverlee, Belgium.

Show MeSH
Related in: MedlinePlus