Thapsigargin blocks <i>Pseudomonas aeruginosa</i> homoserine lactone-induced apoptosis in airway epithelia

Schwarzer, Christian; Ravishankar, B.; Patanwala, Maria; Shuai, Stacey; Fu, Zhu; Illek, Beate; Fischer, Horst; Machen, Terry E.

doi:10.1152/ajpcell.00002.2014

Cited by 25 publications

(16 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Propidium iodide (1 μM) was included to identify dead epithelial cells as indicated in the figure legends. The imaging microscopes used for these experiments have been described previously [ 53 , 54 ].…”

Section: Methodsmentioning

confidence: 99%

Chemotaxis and Binding of Pseudomonas aeruginosa to Scratch-Wounded Human Cystic Fibrosis Airway Epithelial Cells

Schwarzer

Fischer²,

Machen

2016

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Confocal imaging was used to characterize interactions of Pseudomonas aeruginosa (PA, expressing GFP or labeled with Syto 11) with CF airway epithelial cells (CFBE41o-, grown as confluent monolayers with unknown polarity on coverglasses) in control conditions and following scratch wounding. Epithelia and PAO1-GFP or PAK-GFP (2 MOI) were incubated with Ringer containing typical extracellular salts, pH and glucose and propidium iodide (PI, to identify dead cells). PAO1 and PAK swam randomly over and did not bind to nonwounded CFBE41o- cells. PA migrated rapidly (began within 20 sec, maximum by 5 mins) and massively (10–80 fold increase, termed “swarming”), but transiently (random swimming after 15 mins), to wounds, particularly near cells that took up PI. Some PA remained immobilized on cells near the wound. PA swam randomly over intact CFBE41o- monolayers and wounded monolayers that had been incubated with medium for 1 hr. Expression of CFTR and altered pH of the media did not affect PA interactions with CFBE41o- wounds. In contrast, PAO1 swarming and immobilization along wounds was abolished in PAO1 (PAO1ΔcheYZABW, no expression of chemotaxis regulatory components cheY, cheZ, cheA, cheB and cheW) and greatly reduced in PAO1 that did not express amino acid receptors pctA, B and C (PAO1ΔpctABC) and in PAO1 incubated in Ringer containing a high concentration of mixed amino acids. Non-piliated PAKΔpilA swarmed normally towards wounded areas but bound infrequently to CFBE41o- cells. In contrast, both swarming and binding of PA to CFBE41o- cells near wounds were prevented in non-flagellated PAKΔfliC. Data are consistent with the idea that (i) PA use amino acid sensor-driven chemotaxis and flagella-driven swimming to swarm to CF airway epithelial cells near wounds and (ii) PA use pili to bind to epithelial cells near wounds.

show abstract

Section: Methodsmentioning

confidence: 99%

Chemotaxis and Binding of Pseudomonas aeruginosa to Scratch-Wounded Human Cystic Fibrosis Airway Epithelial Cells

Schwarzer

Fischer²,

Machen

2016

PLoS ONE

Self Cite

View full text Add to dashboard Cite

show abstract

“…C12 also triggers apoptosis in multiple cell types (9,(17)(18)(19)(20)(21). That C12 activates apoptosis and not some other type of cell death has been documented from the multiple responses that are classically attributed to apoptosis: membrane blebbing, nuclear condensation and fragmentation, depolarization of mitochondrial membrane potential (⌬ mito ), release of cytochrome c from mitochondria into the cytosol, and activation of caspases 3/7, 8, and 9 and block by the pan-caspase inhibitor Z-VAD-fmk (9,22). A unique aspect of C12-triggered apoptosis was that it occurred equally well in fibroblasts from wild type mouse embryos and from Bax/Bak double knock-out (Bax Ϫ/Ϫ /Bak Ϫ/Ϫ ) mouse embryos (DKO MEF) (23).…”

mentioning

confidence: 99%

Paraoxonase 2 Serves a Proapopotic Function in Mouse and Human Cells in Response to the Pseudomonas aeruginosa Quorum-sensing Molecule N-(3-Oxododecanoyl)-homoserine Lactone

Schwarzer

Morita

et al. 2015

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Background: Pseudomonas aeruginosa use N-(3-oxododecanoyl)-homoserine lactone (C12) for intercellular communication; paraoxonase 2 (PON2) is an animal enzyme that cleaves lactones. Results: C12 elicited apoptosis in mouse and human cells expressing PON2 with intact lactonase activity but not in cells without it. Conclusion: PON2 mediates C12-induced apoptosis in mammalian cells. Significance: PON2 in host cells hydrolyzes C12 and promotes C12-induced apoptosis.

show abstract

“…Exposure to 3O-C12-HSL induces a decrease in the cellular contents and tyrosine-hyperphosphorylation of ZO-1, ZO-3 and JAM-A, and simultaneously triggers occludin dephosphorylation (Vikström et al, 2009 , 2010 ). These modifications lead to a loss of interaction between TJ components, resulting in the transient loss of TJ sealing capacity (Vikström et al, 2009 ; Halldorsson et al, 2010 ; Schwarzer et al, 2014 ). Incubation of epithelial cells with 3O-C12-HSL also decreases cellular levels of E-cadherin/β-catenin and modifies their phosphorylation status, diminishing homophilic binding (Vikström et al, 2009 ).…”

Section: Bacterial Virulence Factors Involved In Junction Disruptionmentioning

confidence: 99%

Pseudomonas aeruginosa Takes a Multi-Target Approach to Achieve Junction Breach

Golovkine

Reboud

Huber

2018

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

show abstract

Thapsigargin blocks Pseudomonas aeruginosa homoserine lactone-induced apoptosis in airway epithelia

Abstract: Machen TE. Thapsigargin blocks Pseudomonas aeruginosa homoserine lactone-induced apoptosis in airway epithelia.

Cited by 25 publications

References 42 publications

Chemotaxis and Binding of Pseudomonas aeruginosa to Scratch-Wounded Human Cystic Fibrosis Airway Epithelial Cells

Chemotaxis and Binding of Pseudomonas aeruginosa to Scratch-Wounded Human Cystic Fibrosis Airway Epithelial Cells

Paraoxonase 2 Serves a Proapopotic Function in Mouse and Human Cells in Response to the Pseudomonas aeruginosa Quorum-sensing Molecule N-(3-Oxododecanoyl)-homoserine Lactone

Pseudomonas aeruginosa Takes a Multi-Target Approach to Achieve Junction Breach

Contact Info

Product

Resources

About