scholarly journals lptGcontributes to changes in membrane permeability and the emergence of multidrug hypersusceptibility in a cystic fibrosis isolate ofPseudomonas aeruginosa

2019 ◽  
Vol 8 (11) ◽  
Author(s):  
Lucas B. Harrison ◽  
Randal C. Fowler ◽  
Baha Abdalhamid ◽  
Anna Selmecki ◽  
Nancy D. Hanson
Keyword(s):  
Cystic Fibrosis ◽  
Membrane Permeability ◽  
Cystic Fibrosis Isolate

scholarly journals Energy-Generating Enzymes of Burkholderia cepacia and Their Interactions with Macrophages

2003 ◽  
Vol 185 (10) ◽  
pp. 3167-3178 ◽  
Author(s):  
Vasu Punj ◽  
Rachna Sharma ◽  
Olga Zaborina ◽  
A. M. Chakrabarty
Keyword(s):  
Cystic Fibrosis ◽  
Cell Death ◽  
Growth Medium ◽  
Burkholderia Cepacia ◽  
Dependent Manner ◽  
Bacterial Protein ◽  
Sepharose Column ◽  
Cellular Energetics ◽  
Cystic Fibrosis Isolate ◽  
High Cytotoxicity

ABSTRACT We previously demonstrated that several clinical and environmental isolates of Burkholderia cepacia secreted ATP-utilizing enzymes to the medium; the secretion of these enzymes by cystic fibrosis lung isolate strain 38 was shown to be greatly enhanced in the presence of α2-macroglobulin. Fractionation of the growth medium of cystic fibrosis isolate strain 71 belonging to genomovar I demonstrated the presence of two additional proteins, homologues of Pseudomonas aeruginosa azurin and cytochrome c 551, which are normally involved in electron transfer during denitrification. A Q-Sepharose column flowthrough fraction of the growth medium of B. cepacia strain 71 enriched with the azurin and cytochrome c 551 homologues triggered apoptosis in macrophages and mast cells, leading to their death. Incubation of the Q-Sepharose column flowthrough fraction with antiazurin and anti-cytochrome c 551 antibodies greatly reduced cell death. We cloned and hyperexpressed a gene from B. cepacia strain 71 that encodes the homologue of P. aeruginosa azurin. Such azurin homologues were detected in the growth medium of several strains belonging to genomovars I, III, and VI but not in the growth medium of strains belonging to other genomovars. The growth medium of the strains that elaborated the azurin homologue had high cytotoxicity towards macrophages. Purified azurin homologue was shown to induce apoptosis in macrophages in a caspase-dependent manner and was localized in both the cytosol and nucleus when incubated with or microinjected into macrophages. This is an interesting example of the interaction of a bacterial protein normally involved in cellular energetics with macrophages to effect their cell death.

scholarly journals Isocitrate Lyase Supplies Precursors for Hydrogen Cyanide Production in a Cystic Fibrosis Isolate of Pseudomonas aeruginosa

2009 ◽  
Vol 191 (20) ◽  
pp. 6335-6339 ◽  
Author(s):  
Jessica M. Hagins ◽  
Robert Locy ◽  
Laura Silo-Suh
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Growth Medium ◽  
Hydrogen Cyanide ◽  
Isocitrate Lyase ◽  
Acid Dehydrogenase ◽  
Amino Acid Dehydrogenase ◽  
Cystic Fibrosis Isolate ◽  
Glyoxylate Pathway

ABSTRACT Pseudomonas aeruginosa colonizes and can persist in the lungs of cystic fibrosis (CF) patients for decades. Adaptation of P. aeruginosa to the CF lung environment causes various genotypic and phenotypic alterations in the bacterium that facilitate persistence. We showed previously that isocitrate lyase (ICL) activity is constitutively upregulated in the P. aeruginosa CF isolate FRD1. We show here that high ICL activity in FRD1 contributes to increased hydrogen cyanide (HCN) production by this isolate. Disruption of aceA, which encodes ICL, results in reduced cyanide production by FRD1 but does not affect cyanide production in the wound isolate PAO1. Cyanide production is restored to the FRD1aceA mutant by addition of glyoxylate, a product of ICL activity, or glycine to the growth medium. Conversion of glyoxylate to glycine may provide a mechanism for increased cyanide production by P. aeruginosa growing on compounds that activate the glyoxylate pathway. Consistent with this hypothesis, disruption of PA5304, encoding a putative d-amino acid dehydrogenase (DadA), led to decreased cyanide production by FRD1. Cyanide production was restored to the FRD1dadA mutant by the addition of glycine, but not glyoxylate, to the growth medium, suggesting that loss of the ability to convert glyoxylate to glycine was associated with the dadA mutation. This was supported by increased glycine production from toluene-treated FRD1 cells with the addition of glyoxylate compared to FRD1dadA cells. This study indicates a larger role for ICL in the physiology and virulence of chronic isolates of P. aeruginosa than previously recognized.

scholarly journals Structural analysis of the lipopolysaccharide core of a rough, cystic fibrosis isolate of Pseudomonas aeruginosa

2001 ◽  
Vol 268 (17) ◽  
pp. 4708-4719 ◽  
Author(s):  
Yuriy A. Knirel ◽  
Ol'ga V. Bystrova ◽  
Alexander S. Shashkov ◽  
Buko Lindner ◽  
Nina A. Kocharova ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Structural Analysis ◽  
Cystic Fibrosis Isolate

scholarly journals Complete Genome Sequence of Persistent Cystic Fibrosis Isolate Pseudomonas aeruginosa Strain RP73

2013 ◽  
Vol 1 (4) ◽  
Author(s):  
J. Jeukens ◽  
B. Boyle ◽  
I. Bianconi ◽  
I. Kukavica-Ibrulj ◽  
B. Tummler ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Cystic Fibrosis Isolate

Malate synthase expression is deregulated in the Pseudomonas aeruginosa cystic fibrosis isolate FRD1

2011 ◽  
Vol 57 (3) ◽  
pp. 186-195 ◽  
Author(s):  
Jessica M. Hagins ◽  
Jessica Scoffield ◽  
Sang-Jin Suh ◽  
Laura Silo-Suh
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Carbon Sources ◽  
Malate Synthase ◽  
Current Evidence ◽  
Pathway Expression ◽  
Genes Encoding ◽  
Key Enzyme ◽  
Cystic Fibrosis Isolate ◽  
Glyoxylate Pathway

Pseudomonas aeruginosa causes chronic pulmonary infections, which can persist for decades, in patients with cystic fibrosis (CF). Current evidence suggests that the glyoxylate pathway is an important metabolic pathway for P. aeruginosa growing within the CF lung. In this study, we identified glcB, which encodes for the second key enzyme of the glyoxylate pathway, malate synthase, as a requirement for virulence of P. aeruginosa on alfalfa seedlings. While expression of glcB in PAO1, an acute isolate of P. aeruginosa, responds to some carbon sources that use the glyoxylate pathway, expression of glcB in FRD1, a CF isolate, is constitutively upregulated. Malate synthase activity is moderately affected by glcB expression and is nearly constitutive in both backgrounds, with slightly higher activity in FRD1 than in PAO1. In addition, RpoN negatively regulates glcB in PAO1 but not in FRD1. In summary, the genes encoding for the glyoxylate-specific enzymes appear to be coordinately regulated, even though they are not located within the same operon on the P. aeruginosa genome. Furthermore, both genes encoding for the glyoxylate enzymes can become deregulated during adaptation of the bacterium to the CF lung.

scholarly journals Adenylate Kinase as a Virulence Factor ofPseudomonas aeruginosa

2001 ◽  
Vol 183 (11) ◽  
pp. 3345-3352 ◽  
Author(s):  
Adam Markaryan ◽  
Olga Zaborina ◽  
Vasu Punj ◽  
A. M. Chakrabarty
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Cell Death ◽  
Virulence Factor ◽  
Adenylate Kinase ◽  
Adenine Nucleotides ◽  
Macrophage Cell ◽  
Cystic Fibrosis Isolate ◽  
Macrophage Killing

ABSTRACT Adenylate kinase (AK; ATP:AMP phosphotransferase, EC 2.7.4.3 ) is a ubiquitous enzyme that contributes to the homeostasis of adenine nucleotides in eukaryotic and prokaryotic cells. AK catalyzes the reversible reaction Mg · ATP + AMP ↔ Mg · ADP + ADP. In this study we show that AK secreted by the pathogenic strains of Pseudomonas aeruginosa appears to play an important role in macrophage cell death. We purified and characterized AK from the growth medium of a cystic fibrosis isolate strain of P. aeruginosa 8821 and hyperproduced it as a fusion protein with glutathione S-transferase. We demonstrated enhanced macrophage cell death in the presence of both the secreted and recombinant purified AK and its substrates AMP plus ATP or ADP. These data suggested that AK converts its substrates to a mixture of AMP, ADP, and ATP, which are potentially more cytotoxic than ATP alone. In addition, we observed increased macrophage killing in the presence of AK and ATP alone. Since the presence of ATPase activity on the macrophages was confirmed in the present work, external macrophage-effluxed ATP is converted to ADP, which in turn can be transformed by AK into a cytotoxic mixture of three adenine nucleotides. Evidence is presented in this study that secreted AK was detected in macrophages during infection with P. aeruginosa. Thus, the possible role of secreted AK as a virulence factor is in producing and keeping an intact pool of toxic mixtures of AMP, ADP, and ATP, which allows P. aeruginosa to exert its full virulence.

Virulence determinants from a cystic fibrosis isolate of Pseudomonas aeruginosa include isocitrate lyase

Microbiology ◽  
2008 ◽  
Vol 154 (6) ◽  
pp. 1616-1627 ◽  
Author(s):  
Tamishia L. Lindsey ◽  
Jessica M. Hagins ◽  
Pamela A. Sokol ◽  
Laura A. Silo-Suh
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Isocitrate Lyase ◽  
Virulence Determinants ◽  
Cystic Fibrosis Isolate
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