Transposon mutations in the flagella biosynthetic pathway of the solvent-tolerant Pseudomonas putida S12 result in a decreased expression of solvent efflux genes

Kieboom, J.

doi:10.1016/s0378-1097(01)00119-7

Cited by 11 publications

(15 citation statements)

References 31 publications

(47 reference statements)

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“…Early studies by Ramos et al (21) revealed that P. putida DOT-T1E with innate solvent tolerance also showed traits related to phase variation; i.e., 1 out of 10 4 cells survived a sudden toluene shock. Our group and other groups had previously found that mutations in the flagellar export machinery led to solvent sensitivity in P. putida DOT-T1E (24) and S12 (13). Interestingly, we have now found in P. putida DOT-T1E a phase variation associated with flagellum biosynthesis linked to the flhB gene, which correlates with the innate solvent tolerance of this strain.…”

supporting

confidence: 66%

“…In Pseudomonas putida DOT-T1E and P. putida S12, flagellar genes have been shown to be involved in organic solvent tolerance, although the role of their proteins in solvent tolerance remains unclear (13,24). Organic solvents with a logP ow (logarithm of its partition coefficient in n-octanol and water) between 1.5 and 4.0 are extremely toxic for microorganisms and other living cells because they partition preferentially in the cytoplasmic membrane, disorganizing its structure and impairing vital functions.…”

mentioning

confidence: 99%

“…Given that mutated FliP, FlhB, and other proteins involved in flagellum export were sensitive to solvents, we hypothesized that the flagellar export machinery could be involved in the possible translocation of toluene tolerance proteins (such as some components of the efflux pumps) to the periplasmic space (13,24).…”

mentioning

confidence: 99%

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Transcriptional Phase Variation at the flhB Gene of Pseudomonas putida DOT-T1E Is Involved in Response to Environmental Changes and Suggests the Participation of the Flagellar Export System in Solvent Tolerance

et al. 2004

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Frameshift mutations in a poly(G) track at the flhB gene of Pseudomonas putida DOT-T1E are responsible for the diminished swimming of this strain on semisolid medium, which contrasts with the high swimming ability of P. putida KT2440, which does not exhibit a poly(G) track at the flhB gene. We previously showed that a mutant lacking FlhB was more sensitive to solvents than the wild-type strain (Segura et al., J. Bacteriol., 183:4127-4133, 2001). In this study, we show that swimming ability correlates with solvent tolerance in P. putida DOT-T1E, so that growth conditions favoring a functional flhB gene (growth on semisolid medium) resulted in increased innate tolerance to a sudden toluene shock.

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confidence: 66%

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confidence: 99%

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Transcriptional Phase Variation at the flhB Gene of Pseudomonas putida DOT-T1E Is Involved in Response to Environmental Changes and Suggests the Participation of the Flagellar Export System in Solvent Tolerance

et al. 2004

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“…However, the C-terminal 100 amino acids of the SigF proteins do not have similarities to other proteins or domains, including the region 4 domains (Pfam domain number PF04545) containing the Ϫ35 promoter-binding element. AY-WB SigF proteins showed the greatest similarity (E value, 10 Ϫ6 ) to the stress response sigma factor [sigma(H)] of Streptococcus coelicolor (50) and the flagellar biosynthesis sigma factor FliA of Pseudomonas putida (46). Expression of SigF and other PMU genes might occur under specific environmental conditions.…”

Section: Discussionmentioning

confidence: 99%

Living with Genome Instability: the Adaptation of Phytoplasmas to Diverse Environments of Their Insect and Plant Hosts

Bai

Zhang

Ewing³

et al. 2006

J Bacteriol

343

447

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Phytoplasmas ("CandidatusPhytoplasmaThe phylogenetic tree of mollicutes is composed of two major clades that diverged early in evolution (51). One clade contains the orders Acholeplasmatales and Anaeroplasmatales (AAA clade mollicutes), and the other clade contains the orders Mycoplasmatales and Entomoplasmatales (SEM clade mollicutes) (9). Phytoplasmas, formerly known as mycoplasma-like organisms of plants, form a monophyletic group in the order Acholeplasmatales (51) and were recently assigned to a novel genus, "Candidatus Phytoplasma" (41). Approximately 20 phytoplasma phylogenetic groups have been proposed based on 16S rRNA gene sequences, and new branches are continuously being discovered (69,85). Members of the order Acholeplasmatales are distinct from other mollicutes in several ways. For instance, whereas most mollicutes use UGA as a tryptophan codon instead of a stop codon, a feature they share with mitochondria, the acholeplasmas and phytoplasmas retained UGA as a stop codon (80).

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“…Although extrusion of the solvent by efflux pumps is probably the most important mechanism leading to solvent resistance, some other factors have been identified as being involved in the solvent tolerance phenomenon. Among these factors are membrane rigidification via isomerization of the cis unsaturated fatty acids into the corresponding trans isomers (18,22), flagellar export proteins (24,50), and formation of membrane vesicles filled with toluene (26).…”

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confidence: 99%

Proteomic Analysis Reveals the Participation of Energy- and Stress-Related Proteins in the Response of Pseudomonas putida DOT-T1E to Toluene

et al. 2005

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Pseudomonas putida DOT-T1E is tolerant to toluene and other toxic hydrocarbons through extrusion of the toxic compounds from the cell by means of three efflux pumps, TtgABC, TtgDEF, and TtgGHI. To identify other cellular factors that allow the growth of P. putida DOT-T1E in the presence of high concentrations of toluene, we performed two-dimensional gel analyses of proteins extracted from cultures grown on glucose in the presence and in the absence of the organic solvent. From a total of 531 spots, 134 proteins were observed to be toluene specific. In the absence of toluene, 525 spots were clearly separated and 117 proteins were only present in this condition. Moreover, 35 proteins were induced by at least twofold in the presence of toluene whereas 26 were repressed by at least twofold under these conditions. We reasoned that proteins that were highly induced could play a role in toluene tolerance. These proteins, identified by matrix-assisted laser desorption ionizationtime of flight mass spectrometry, were classified into four categories: 1, proteins involved in the catabolism of toluene; 2, proteins involved in the channeling of metabolic intermediates to the Krebs cycle and activation of purine biosynthesis; 3, proteins involved in sugar transport; 4, stress-related proteins. The set of proteins in groups 2 and 3 suggests that the high energy demand required for solvent tolerance is achieved via activation of cell metabolism. The role of chaperones that facilitate the proper folding of newly synthesized proteins under toluene stress conditions was analyzed in further detail. Knockout mutants revealed that CspA, XenA, and Tuf-1 play a role in solvent tolerance in Pseudomonas, although this role is probably not specific to toluene, as indicated by the fact that all mutants grew more slowly than the wild type without toluene.Organic solvents with a log P ow (logarithm of the partition coefficient in a mixture of octanol and water) between 1.5 and 4 are extremely toxic for microorganisms because they partition in the cell membranes and disorganize them by removing lipids and proteins, eventually leading to cell death (8). However, since the report by Inoue and Horikoshi (19) of a bacterium able to thrive in the presence of high concentrations of organic solvents, other solvent-tolerant bacteria have been isolated. Among them, Pseudomonas putida DOT-T1E, a highly solvent-tolerant microbe, was isolated from a wastewater treatment plant in Granada, Spain (40). Solvent tolerance in P. putida DOT-T1E is an inducible phenomenon. When cells were shocked with 0.3% (vol/vol) toluene, only 1 out of 10,000 survived; however, if cells were preinduced with a small amount of toluene (i.e., 0.01% [vol/vol]), almost 100% of the cells survived the 0.3% (vol/vol) toluene shock.The mechanisms underlying solvent tolerance are not yet fully understood, but a number of factors involved in the process have been characterized over the last 10 years (43). Several laboratories have identified efflux pumps belonging to the RND (resistance...

show abstract

Transposon mutations in the flagella biosynthetic pathway of the solvent-tolerant Pseudomonas putida S12 result in a decreased expression of solvent efflux genes

Cited by 11 publications

References 31 publications

Transcriptional Phase Variation at the flhB Gene of Pseudomonas putida DOT-T1E Is Involved in Response to Environmental Changes and Suggests the Participation of the Flagellar Export System in Solvent Tolerance

Transcriptional Phase Variation at the flhB Gene of Pseudomonas putida DOT-T1E Is Involved in Response to Environmental Changes and Suggests the Participation of the Flagellar Export System in Solvent Tolerance

Living with Genome Instability: the Adaptation of Phytoplasmas to Diverse Environments of Their Insect and Plant Hosts

Proteomic Analysis Reveals the Participation of Energy- and Stress-Related Proteins in the Response of Pseudomonas putida DOT-T1E to Toluene

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