Cloning and nucleotide sequence of the <i>Pseudomonas aeruginosa</i> glucose‐selective OprB porin gene and distribution of OprB within the family <i>Pseudomonadaceae</i>

Wylie, John; Worobec, Elizabeth A.

doi:10.1111/j.1432-1033.1994.tb18649.x

Cited by 29 publications

(16 citation statements)

References 35 publications

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“…The cloning and nucleotide sequence of the P. aeruginosa oprB gene have been previously described (36). The allele replacement technique of Schweizer (26) was used to construct a defined OprB-lacking mutant strain.…”

Section: Resultsmentioning

confidence: 99%

The OprB porin plays a central role in carbohydrate uptake in Pseudomonas aeruginosa

Wylie

Worobec

1995

J Bacteriol

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Using interposon mutagenesis, we have generated strains of Pseudomonas aeruginosa which lack or overexpress the substrate-selective OprB porin of this species. A marked decrease or increase in the initial uptake of glucose by these strains verified the role of OprB in facilitating the diffusion of glucose across the outer membrane of P. aeruginosa. However, we also demonstrated that the loss or overexpression of OprB had a similar effect on the uptake of three other sugars able to support the growth of this bacterium (mannitol, glycerol, and fructose). This effect was restricted to carbohydrate transport; arginine uptake was identical in mutant and wild-type strains. These results indicated that OprB cannot be considered strictly a glucoseselective porin; rather, it acts as a central component of carbohydrate transport and is more accurately described as a carbohydrate-selective porin.Diffusion rates across the outer membrane of Pseudomonas aeruginosa are 100 to 500 times lower than those measured for Escherichia coli (37). The low outer membrane permeability of P. aeruginosa has been attributed to the unusual channel-forming properties of the OprF porin, the major, constitutively expressed protein in the outer membrane of this bacterium (9). OprF has some homology with the OmpA protein of enteric bacteria (33) and, like OmpA, shows only limited porin activity (28). In enteric bacteria, the low channel-forming activity of OmpA does not have a major influence on outer membrane permeability, as these bacteria also produce classical general diffusion porins (e.g., OmpF and OmpC). The more efficient permeation of solutes through OmpF and OmpC overshadows the diffusion properties of the OmpA protein. In contrast, the apparent lack of typical general diffusion porins in P. aeruginosa results in the diffusion properties of the outer membrane of this bacterium being determined largely by the relatively inefficient OprF porin.The low outer membrane permeability of P. aeruginosa is compensated for by the presence of several substrate-selective porins (14,21,29). Porins of this type are also present in enteric bacteria, but in these species, substrate-selective porins have generally been associated with the diffusion of relatively large substrates (e.g., LamB, maltodextrins [32]; ScrY, sucrose [25]; TolC, peptides [2]). In the absence of their respective porins, these substrates either do not diffuse or diffuse to only a limited extent, through nonspecific channels. In contrast, the substrate-selective porins of P. aeruginosa facilitate the diffusion of much smaller substrates. In E. coli, diffusion of small substrates, such as glucose or glycerol, is generally believed to occur via nonspecific channels. In P. aeruginosa, if facilitated diffusion across the outer membrane did not occur, the low outer membrane permeability would potentially limit the transport of almost all substrates, regardless of size.Four substrate-selective porins have been identified in P. aeruginosa. The OprD porin facilitates the diffusion of basic ...

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Section: Resultsmentioning

confidence: 99%

The OprB porin plays a central role in carbohydrate uptake in Pseudomonas aeruginosa

Wylie

Worobec

1995

J Bacteriol

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“…Therefore, it is possible that one of the many genes encoding hypothetical proteins identified in this study encodes one of these two enzymes. Also a sugar ABC transporter gene cluster (PP1015-PP1019; Wylie & Worobec, 1994, 1995 was identified that was specifically induced upon growth on glucose (Table 3). This gene cluster encodes a glucose porin and a sugar transporter, of which the sugarbinding protein (PP1015) is very likely the previously purified glucose-specific glucose binding protein (Stinson et al, 1977), as this gene encodes a protein of a similar size as the purified protein (44?5 kDa) and has a similar amino acid composition.…”

Section: Hcamentioning

confidence: 99%

Multivariate analysis of microarray data by principal component discriminant analysis: prioritizing relevant transcripts linked to the degradation of different carbohydrates in Pseudomonas putida S12

et al. 2006

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The value of the multivariate data analysis tools principal component analysis (PCA) and principal component discriminant analysis (PCDA) for prioritizing leads generated by microarrays was evaluated. To this end, Pseudomonas putida S12 was grown in independent triplicate fermentations on four different carbon sources, i.e. fructose, glucose, gluconate and succinate. RNA isolated from these samples was analysed in duplicate on an anonymous clone-based array to avoid bias during data analysis. The relevant transcripts were identified by analysing the loadings of the principal components (PC) and discriminants (D) in PCA and PCDA, respectively. Even more specifically, the relevant transcripts for a specific phenotype could also be ranked from the loadings under an angle (biplot) obtained after PCDA analysis. The leads identified in this way were compared with those identified using the commonly applied fold-difference and hierarchical clustering approaches. The different data analysis methods gave different results. The methods used were complementary and together resulted in a comprehensive picture of the processes important for the different carbon sources studied. For the more subtle, regulatory processes in a cell, the PCDA approach seemed to be the most effective. Except for glucose and gluconate dehydrogenase, all genes involved in the degradation of glucose, gluconate and fructose were identified. Moreover, the transcriptomics approach resulted in potential new insights into the physiology of the degradation of these carbon sources. Indications of iron limitation were observed with cells grown on glucose, gluconate or succinate but not with fructose-grown cells. Moreover, several cytochrome- or quinone-associated genes seemed to be specifically up- or downregulated, indicating that the composition of the electron-transport chain in P. putida S12 might change significantly in fructose-grown cells compared to glucose-, gluconate- or succinate-grown cells.

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“…The full complement of genes and protein components of this system has not been described. Presumably, two of the genes regulated by GltR are oprD, which encodes a glucose-specific porin (23,24), and the yet to be identified gene for GBP (gltB). The OmpR subfamily of these proteins control a variety of systems in different bacteria, including those involved in response to medium osmolarity, phosphate uptake, virulence, and antibiotic resistance (12).…”

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confidence: 99%

A two-component response regulator, gltR, is required for glucose transport activity in Pseudomonas aeruginosa PAO1

1996

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A 729-bp open reading frame (gltR) was identified in Pseudomonas aeruginosa PAO1 that encodes a product homologous to the two-component response regulator family of proteins. Disruption of gltR caused loss of glucose transport activity. Restoration of gltR resulted in wild-type levels of glucose transport. These findings indicate that gltR is required for expression of the glucose transport system in P. aeruginosa.

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Cloning and nucleotide sequence of the Pseudomonas aeruginosa glucose‐selective OprB porin gene and distribution of OprB within the family Pseudomonadaceae

Cited by 29 publications

References 35 publications

The OprB porin plays a central role in carbohydrate uptake in Pseudomonas aeruginosa

The OprB porin plays a central role in carbohydrate uptake in Pseudomonas aeruginosa

Multivariate analysis of microarray data by principal component discriminant analysis: prioritizing relevant transcripts linked to the degradation of different carbohydrates in Pseudomonas putida S12

A two-component response regulator, gltR, is required for glucose transport activity in Pseudomonas aeruginosa PAO1

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