The clonal antibody response to <i>Pseudomonas aeruginosa</i> heat shock protein is highly diverse in cystic fibrosis patients

Ulanova, Marina; Petersen, T. D.; Ciofu, Oana; Jensen, Peter Kjær Mackie; Hahn-Zoric, Mirjana; Hanson, Lars Å.; Høiby, Niels

doi:10.1111/j.1699-0463.1997.tb00593.x

Cited by 7 publications

(4 citation statements)

References 22 publications

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“…GroES, HtpG, and GroEL were significantly downregulated. Previous work has shown that CF patients produce antibodies to GroEL (Ulanova et al, 1997 ). Other recent work has shown that heat shock proteins such as GroES and HtpG are actually downregulated in biofilms (Patrauchan et al, 2007 ).…”

Section: Resultsmentioning

confidence: 98%

Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient

Rao¹,

Damron²,

Basler³

et al. 2011

Front. Microbio.

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Pseudomonas aeruginosa chronically infects the lungs of cystic fibrosis (CF) patients. The conditions in the CF lung appear to select for P. aeruginosa with advantageous phenotypes for chronic infection. However, the mechanisms that allow the establishment of this chronic infection have not been fully characterized. We have previously reported the transcriptional analysis of two CF isolates strains 383 and 2192. Strain 2192 is a mucoid, alginate overproducing strain whereas strain 383 is non-mucoid. Mucoid strains are associated with chronic infection of the CF lung and non-mucoid strains are the typical initially infecting isolates. To elucidate novel differences between these two strains, we employed two methods of shotgun proteomics: isobaric tags for relative and absolute quantitation (iTRAQ) and two-dimensional gel electrophoresis (2-DE). iTRAQ compares the amount of protein between samples and relies on protein abundance, while 2-DE gel electrophoresis depends on selection of separated protein spots. For both these methods, mass spectrometry was then used to identify proteins differentially expressed between the two strains. The compilation of these two proteomic methods along with Western blot analysis revealed proteins of the HSI-I operon of the type 6 secretion system, showed increased expression in 383 compared to 2192, confirming the our previous transcriptional analysis. Proteomic analysis of other proteins did not fully correlate with the transcriptome but other differentially expressed proteins are discussed. Also, differences were noted between the results obtained for the two proteomic techniques. These shotgun proteomic analyses identified proteins that had been predicted only through gene identification; we now refer to these as “proteins of unknown functions” since their existence has now been established however their functional characterization remains to be elucidated.

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Section: Resultsmentioning

confidence: 98%

Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient

Rao¹,

Damron²,

Basler³

et al. 2011

Front. Microbio.

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“…This suggests PIA‐AMV may cause protein misfolding, which would cause upregulation of chaperones to maintain the cellular integrity. Previously, it has been observed that chronically infected CF patients have a high antibody titre against GroEL (Ulanova et al ., 1997). Since PIA‐AMV seems to upregulate GroEL (Table 2), this further suggests PIA‐AMV mimics the CF lung condition.…”

Section: Resultsmentioning

confidence: 99%

Vanadate and triclosan synergistically induce alginate production by Pseudomonas aeruginosa strain PAO1

Damron

Davis

Withers³

et al. 2011

Molecular Microbiology

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Summary Alginate overproduction by P. aeruginosa strains, also known as mucoidy, is associated with chronic lung infections in cystic fibrosis (CF). It is not clear how alginate induction occurs in the wild type (wt) mucA strains. When grown on Pseudomonas isolation agar (PIA), P. aeruginosa strains PAO1 and PA14 are nonmucoid producing minimal amounts of alginate. Here we report the addition of ammonium metavanadate (AMV), a phosphatase inhibitor, to PIA (PIA-AMV) induced mucoidy in both these laboratory strains and early lung colonizing nonmucoid isolates with a wt mucA. This phenotypic switch was reversible depending on the availability of vanadate salts and triclosan, a component of PIA. Alginate induction in PAO1 on PIA-AMV was correlated with increased proteolytic degradation of MucA, and required envelope proteases AlgW or MucP, and a two-component phosphate regulator, PhoP. Other changes included the addition of palmitate to lipid A, a phenotype also observed in chronic CF isolates. Proteomic analysis revealed the upregulation of stress chaperones, which was confirmed by increased expression of the chaperone/protease MucD. Altogether, these findings suggest a model of alginate induction and the PIA-AMV medium may be suitable for examining early lung colonization phenotypes in CF before the selection of the mucA mutants.

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“…GroEL is among the most highly conserved proteins in nature (Segal & Ron, 1996), and functions together with GroES to maintain protein integrity, which enables cells to survive a variety of environmental stresses (Hendrick & Hartl, 1993). A strong antibody response to GroEL has been found in cystic fibrosis patients with chronic pulmonary infection caused by P. aeruginosa (Ulanova et al, 1997), which suggests a role for PhrS under these conditions. The outer membrane porin OprD has been shown to facilitate diffusion of basic amino acids as well as of small peptides, and also serves as a channel for the b-lactam antibiotic imipenem (Trias & Nikaido 1990), whereby the loss of OprD can lead to imipenem resistance (Lynch et al, 1987).…”

Section: Proteome Analyses Upon Overproduction Of Phrsmentioning

confidence: 99%

Detection of small RNAs in Pseudomonas aeruginosa by RNomics and structure-based bioinformatic tools

et al. 2008

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Inactivation of the Pseudomonas aeruginosa (PAO1) hfq gene, encoding the Sm-like Hfq protein, resulted in pleiotropic effects that included an attenuated virulence. As regulation by Hfq often involves the action of small regulatory RNAs (sRNAs), we have used a shotgun cloning approach (RNomics) and bioinformatic tools to identify sRNAs in strain PAO1. For cDNA library construction, total RNA was extracted from PAO1 cultures either grown to stationary phase or exposed to human serum. The cDNA libraries were generated from small-sized RNAs of PAO1 after co-immunoprecipitation with Hfq. Of 400 sequenced cDNA clones, 11 mapped to intergenic regions. Band-shift assays and Northern blot analyses performed with two selected sRNAs confirmed that Hfq binds to and affects the steady-state levels of these RNAs. A proteome study performed upon overproduction of one sRNA, PhrS, implicated it in riboregulation. PhrS contains an ORF, and evidence for its translation is presented. In addition, based on surveys with structure-based bioinformatic tools, we provide an electronic compilation of putative sRNA and non-coding RNA genes of PAO1 based on their evolutionarily conserved structure.

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The clonal antibody response to Pseudomonas aeruginosa heat shock protein is highly diverse in cystic fibrosis patients

Cited by 7 publications

References 22 publications

Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient

Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient

Vanadate and triclosan synergistically induce alginate production by Pseudomonas aeruginosa strain PAO1

Detection of small RNAs in Pseudomonas aeruginosa by RNomics and structure-based bioinformatic tools

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