Genetic Characterization of a Multicomponent Signal Transduction System Controlling the Expression of Cable Pili in
            <i>Burkholderia cenocepacia</i>

Tomich, Mladen; Mohr, Christian

doi:10.1128/jb.186.12.3826-3836.2004

Cited by 14 publications

(13 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2). To determine whether expression of the adhesin is regulated by genes that control the expression of Cbl pili, we created an isogenic mutant of cblS, one of the regulatory genes of a multicomponent regulatory system that regulates the biogenesis of Cbl pili (48). Successful insertion of the trimethoprim resistance cassette in all three mutants was confirmed by PCR and Southern blot analysis (data not shown).…”

Section: Identification Of the Gene (Adha) Encoding The 22-kda Ad-mentioning

confidence: 99%

“…The first four genes in the operon, cblB, cblA, cblC, and cblD, are sufficient for pilus biogenesis, as determined by heterologous expression in Escherichia coli (32). However, the regulatory genes cblS, cblT, and cblR are necessary for pilus biogenesis in B. cenocepacia (48). Although the organization of genes necessary for pilus biogenesis in the cbl operon and their predicted products are very similar to enterotoxigenic E. coli CS and CFA/I families (33,35), they differ from the CS and CFA/I class of pili with regard to the adhesin.…”

mentioning

confidence: 99%

“…The cbl operon consists of at least seven genes: cblB, a proposed chaperone-like protein; cblA, the major pilin subunit; cblC, a proposed usher protein; cblD, a minor pilin protein; and the regulatory genes cblS, cblT, and cblR (32,48). The first four genes in the operon, cblB, cblA, cblC, and cblD, are sufficient for pilus biogenesis, as determined by heterologous expression in Escherichia coli (32).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Cable Pili and the 22-Kilodalton Adhesin Are Required for Burkholderia cenocepacia Binding to and Transmigration across the Squamous Epithelium

et al. 2005

View full text Add to dashboard Cite

Burkholderia cenocepacia strains expressing both cable (Cbl) pili and the 22-kDa adhesin bind to cytokeratin 13 (CK13) strongly and invade squamous epithelium efficiently. It has not been established, however, whether the gene encoding the adhesin is located in the cbl operon or what specific contribution the adhesin and Cbl pili lend to binding and transmigration or invasion capacity of B. cenocepacia. By immunoscreening an expression library of B. cenocepacia isolate BC7, we identified a large gene (adhA) that encodes the 22-kDa adhesin. Isogenic mutants lacking expression of either Cbl pili (cblA or cblS mutants) or the adhesin (adhA mutant) were constructed to assess the individual role of Cbl pili and the adhesin in mediating B. cenocepacia binding to and transmigration across squamous epithelium. Relative to the parent strain, mutants of Cbl pili showed reduced binding (50%) to isolated CK13, while the adhesin mutant showed almost no binding (0 to 8%). Mutants lacking either cable pili or the adhesin were compromised in their ability to bind to and transmigrate across the squamous epithelium compared to the wild-type strain, although this deficiency was most pronounced in the adhA mutant. These results indicate that both Cbl pili and the 22-kDa adhesin are necessary for the optimal binding to CK13 and transmigration properties of B. cenocepacia.

show abstract

Section: Identification Of the Gene (Adha) Encoding The 22-kda Ad-mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Cable Pili and the 22-Kilodalton Adhesin Are Required for Burkholderia cenocepacia Binding to and Transmigration across the Squamous Epithelium

et al. 2005

View full text Add to dashboard Cite

show abstract

“…1A). The cblD gene is transcribed as part of the cbl operon, which comprises cblBACDS (19,24). This operon is adjacent to the separately transcribed genes-cblR, which encodes a two-component regulator, and cblT, which encodes a sensor histidine kinase.…”

mentioning

confidence: 99%

Intraspecies Signaling Involving the Diffusible Signal Factor BDSF (cis-2-Dodecenoic Acid) Influences Virulence inBurkholderia cenocepacia

et al. 2009

View full text Add to dashboard Cite

Burkholderia cenocepacia produces a diffusible fatty acid signal molecule, cis-2-dodecenoic acid (BDSF), that has been implicated in interspecies and interkingdom communication. Here, we show that BDSF also acts as an intraspecies signal in B. cenocepacia to control factors contributing to virulence of this major opportunistic pathogen.Many bacteria use cell-cell communication systems involving the synthesis and perception of diffusible signal molecules to monitor aspects of their environment such as cell density or confinement to niches and to modulate their behavior accordingly (reviewed in references 3, 7, 16, 26, and 27). The signal molecules produced by bacteria are structurally diverse. Many gram-negative bacteria use N-acyl homoserine lactones (NAHLs) as signals, although other fatty acid derivatives, including cis-unsaturated fatty acids, are also found. The first example of this latter class of molecule to be described was the diffusible signal factor DSF from the plant pathogen Xanthomonas campestris pv. campestris, which is cis-11-methyl-2-dodecenoic acid (1, 25). Until recently, it was thought that signal molecules of the DSF family were restricted to the xanthomonads, which do not synthesize N-AHLs (4, 11). However, work by Boon and colleagues (2) has demonstrated that Burkholderia cenocepacia, the opportunistic pathogen of cystic fibrosis patients and immunocompromised individuals, produces the DSF-like molecule cis-2-dodecenoic acid (BDSF). This molecule is able to activate DSF-dependent responses in X. campestris pv. campestris and to inhibit germ tube formation in Candida albicans, suggesting a role in interspecies and interkingdom communication. B. cenocepacia produces N-AHLs to regulate a wide range of functions that include virulence, biofilm formation, and motility (9, 23). This raises the question as to whether BDSF has any role as an intraspecies signal in B. cenocepacia J2315, an issue that we address here.BDSF synthesis requires BCAM0581, which is related to the DSF synthase RpfF of X. campestris pv. campestris (1, 2). We have examined the role of BDSF in signaling by comparing the phenotypes of wild-type B. cenocepacia J2315, an rpfF mutant with a deletion in bcam0581, and a complemented mutant strain (Table 1). A strain carrying an in-frame deletion of rpfF was constructed by allelic exchange. DNA fragments comprising the upstream and downstream regions flanking rpfF in the genome were amplified by PCR, using B. cenocepacia genomic DNA as a template and the primer sets RPFFUF/RPFFUR and RPFFDF/ RPFFDR, respectively (Table 1). The amplicons were mixed, diluted, and used as a template for PCR with RPFFUF and RPFFDR as the primers. To facilitate construction, restriction sites for BamHI and HindIII were engineered into these primer sequences. The amplified DNA fragment was cut with BamHI and HindIII and ligated into the allelic exchange vector pEX18Tc to give pEXRPFF. The construction was verified by DNA sequencing. The construct was introduced into B. cenocepacia J2315 by electroporation...

show abstract

“…Standard electrotransformation techniques are ineffective with this strain, as also found elsewhere (26). Conjugal introduction of DNA has proved unreliable despite adaptations (7) that have enabled occasional successes with B. cenocepacia species (9,40) including J2315 (39) (see also Results below). Besides, the natural resistance of J2315 to antibiotics, high even on the scale of the generally extensive resistance of B. cenocepacia species (31), severely restricts the use of antibiotic resistance in genetic selections.…”

mentioning

confidence: 99%

Improved Electrotransformation and Decreased Antibiotic Resistance of the Cystic Fibrosis Pathogen Burkholderia cenocepacia Strain J2315

Dubarry

Lane

et al. 2010

Appl Environ Microbiol

View full text Add to dashboard Cite

The bacterium Burkholderia cenocepacia is pathogenic for sufferers from cystic fibrosis (CF) and certain immunocompromised conditions. The B. cenocepacia strain most frequently isolated from CF patients, and which serves as the reference for CF epidemiology, is J2315. The J2315 genome is split into three chromosomes and one plasmid. The strain was sequenced several years ago, and its annotation has been released recently. This information should allow genetic experimentation with J2315, but two major impediments appear: the poor potential of J2315 to act as a recipient in transformation and conjugation and the high level of resistance it mounts to nearly all antibiotics. Here, we describe modifications to the standard electroporation procedure that allow routine transformation of J2315 by DNA. In addition, we show that deletion of an efflux pump gene and addition of spermine to the medium enhance the sensitivity of J2315 to certain commonly used antibiotics and so allow a wider range of antibiotic resistance genes to be used for selection.

show abstract

Genetic Characterization of a Multicomponent Signal Transduction System Controlling the Expression of Cable Pili in Burkholderia cenocepacia

Cited by 14 publications

References 45 publications

Cable Pili and the 22-Kilodalton Adhesin Are Required for Burkholderia cenocepacia Binding to and Transmigration across the Squamous Epithelium

Cable Pili and the 22-Kilodalton Adhesin Are Required for Burkholderia cenocepacia Binding to and Transmigration across the Squamous Epithelium

Intraspecies Signaling Involving the Diffusible Signal Factor BDSF (cis-2-Dodecenoic Acid) Influences Virulence inBurkholderia cenocepacia

Improved Electrotransformation and Decreased Antibiotic Resistance of the Cystic Fibrosis Pathogen Burkholderia cenocepacia Strain J2315

Contact Info

Product

Resources

About