Exposure to <i>Bordetella pertussis</i> adenylate cyclase toxin affects integrin‐mediated adhesion and mechanics in alveolar epithelial cells

Angely, Christelle; Nguyen, Ngoc-Minh; Dias, Sofia André; Planus, Emmanuelle; Pelle, Gabriel; Louis, Bruno; Filoche, Marcel; Chenal, Alexandre; Ladant, Daniel; Isabey, Daniel

doi:10.1111/boc.201600082

Cited by 9 publications

(17 citation statements)

References 47 publications

(81 reference statements)

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“…287 It has already been shown that CyaA invasion results in a deep remodeling of the actinic 288 CSK localized essentially under the plasma membrane [9,44]. Present data obtained on cellular 289 adhesion as well as previous data obtained on early adhesion in the same cellular system [16] 290 show that CyaA significantly weakens the adhesion system, suggesting that 291 mechanotransduction signaling pathways are affected during the intoxication process.…”

Section: Discussion and Perspectives 275supporting

confidence: 58%

“…Moreover, it has been shown that cAMP 343 modulates cell morphology by inhibiting a Rac-dependent signaling pathway [55]. Similarly, the 344 results obtained in our previous study on the early development of adhesion while the same cells 345 were intoxicated with the same toxin [16], have shown a reduction in chemical energy of 346 individual integrin-RGD ligand bonds as well as a diminution by two of integrin association at 347 the level of newly formed adhesion sites (clustering). For these two processes, the role of Rac 1 348 is fundamental [62] and its potential defect in parallel to the activation of RhoA could be the 349 common cause of these results.…”

mentioning

confidence: 66%

“…Eby et al reported that polarized T84 cell monolayers 27 as well as human airway epithelial cultures could respond to nanomolar concentrations of CyaA 28 when it was applied to the basolateral membranes [15]. We then showed that CyaA can invade 29 A549 alveolar epithelial cells and trigger a significant remodeling of their molecular adhesion 30 systems [16] and, more recently, Hasan et al (2018) analyzed the impact of CyaA on the 31 functional integrity of human bronchial epithelial cells cultured at the air-liquid interface [17]. 32 Cytoskeletal (CSK) rearrangement may be caused by different signal events including 33 modifications in the intracellular Ca 2+ and cAMP-mediated signaling which have been involved 34 in perturbations of the actin CSK homeostasis in different cells [18,19].…”

mentioning

confidence: 76%

“…In our previous study on the early development of adhesion sites in intoxicated cells [16], 293 CyaA was reported to reinforce adhesion by decreasing the strength of newly formed integrin 294 bonds as well as their mutual association through clustering. Thus, alteration of adhesion 295 process and weakening of cell-matrix attachments can be considered as the hallmark of initial 296 January 31, 2020 11/18…”

mentioning

confidence: 97%

“…We previously showed [16] that A549 alveolar epithelial cells are sensitive to CyaA that triggers 191 a cAMP-dependent remodeling of their molecular adhesion system, resulting in a significant the reorganization of the actin cytoskeleton and evolution of focal adhesions. A549 cells were 194 exposed for 1 hr to various concentrations of CyaA (ranging form 0.5 to 10 nM) and then the 195 F-actin structures were visualized by phalloidin staining (in red) and focal adhesions were 196 visualized with anti-phosphotyrosin PY99 antibodies (in green) (Fig 1).…”

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confidence: 99%

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Functional and structural consequences of epithelial cell invasion byBordetella pertussisadenylate cyclase toxin

Angely

Ladant

Planus

et al. 2020

Preprint

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AbstractBordetella pertussis, the causative agent of whopping cough, produces an adenylate cyclase toxin (CyaA) that plays a key role in the host colonization by targeting innate immune cells which express CD11b/CD18, the cellular receptor of CyaA. CyaA is also able to invade non-phagocytic cells, via a unique entry pathway consisting in a direct translocation of its catalytic domain across the cytoplasmic membrane of the cells. Within the cells, CyaA is activated by calmodulin to produce high levels of cyclic adenosine monophosphate (cAMP) and alter cellular physiology. In this study, we explored the effects of CyaA toxin on the cellular and molecular structure remodeling of A549 alveolar epithelial cells. Using classical imaging techniques, biochemical and functional tests, as well as advanced cell mechanics method, we quantify the structural and functional consequences of the massive increase of intracellular cyclic AMP induced by the toxin: cell shape rounding associated to adhesion weakening process, actin structure remodeling for the cortical and dense components, increase in cytoskeleton stiffness, and inhibition of migration and repair. We also show that, at the low concentrations that may be found in vivo during B. pertussis infection, CyaA impairs the migration and wound healing capacities of the intoxicated alveolar epithelial cells. Our results suggest that the CyaA, beyond its major role in disabling innate immune cells, might also contribute to the local alteration of the epithelial barrier of the respiratory tract, that is an hallmark of pertussis.

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Section: Discussion and Perspectives 275supporting

confidence: 58%

mentioning

confidence: 66%

mentioning

confidence: 76%

mentioning

confidence: 97%

mentioning

confidence: 99%

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Functional and structural consequences of epithelial cell invasion byBordetella pertussisadenylate cyclase toxin

Angely

Ladant

Planus

et al. 2020

Preprint

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A High‐Affinity Calmodulin‐Binding Site in the CyaA Toxin Translocation Domain is Essential for Invasion of Eukaryotic Cells

et al. 2021

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The molecular mechanisms and forces involved in the translocation of bacterial toxins into host cells are still a matter of intense research. The adenylate cyclase (CyaA) toxin from Bordetella pertussis displays a unique intoxication pathway in which its catalytic domain is directly translocated across target cell membranes. The CyaA translocation region contains a segment, P454 (residues 454–484), which exhibits membrane‐active properties related to antimicrobial peptides. Herein, the results show that this peptide is able to translocate across membranes and to interact with calmodulin (CaM). Structural and biophysical analyses reveal the key residues of P454 involved in membrane destabilization and calmodulin binding. Mutational analysis demonstrates that these residues play a crucial role in CyaA translocation into target cells. In addition, calmidazolium, a calmodulin inhibitor, efficiently blocks CyaA internalization. It is proposed that after CyaA binding to target cells, the P454 segment destabilizes the plasma membrane, translocates across the lipid bilayer and binds calmodulin. Trapping of CyaA by the CaM:P454 interaction in the cytosol may assist the entry of the N‐terminal catalytic domain by converting the stochastic motion of the polypeptide chain through the membrane into an efficient vectorial chain translocation into host cells.

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Biophysical and biomechanical properties of neural progenitor cells as indicators of developmental neurotoxicity

2019

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Conventional in vitro toxicity studies have focused on identifying IC 50 and the underlying mechanisms, but how toxicants influence biophysical and biomechanical changes in human cells, especially during developmental stages, remain under-studied. Here, using an atomic force microscope, we characterized changes in biophysical (cell area, actin organization) and biomechanical (Young's modulus, force of adhesion, tether force, membrane tension, tether radius) aspects of human fetal brain-derived neural progenitor cells (NPCs) induced by four classes of widely used toxic compounds, including rotenone, digoxin, N-arachidonoylethanolamide (AEA), and chlorpyrifos, under exposure up to 36 h. The sub-cellular mechanisms (apoptosis, mitochondria membrane potential, DNA damage, glutathione levels) by which these toxicants induced biochemical changes in NPCs were assessed. Results suggest a significant compromise in cell viability with increasing toxicant concentration (p < 0.01), and biophysical and biomechanical characteristics with increasing exposure time (p < 0.01) as well as toxicant concentration (p < 0.01). Impairment of mitochondrial membrane potential appears to be the most sensitive mechanism of neurotoxicity for rotenone, AEA and chlorpyrifos exposure, but compromise in plasma membrane integrity for digoxin exposure. The surviving NPCs remarkably retained stemness (SOX2 expression) even at high toxicant concentrations. A negative linear correlation (R 2 = 0.92) exists between the elastic modulus of surviving cells and the number of living cells in that environment. We propose that even subtle compromise in cell mechanics could serve as a crucial marker of developmental neurotoxicity (mechanotoxicology) and therefore should be included as part of toxicology assessment repertoire to characterize as well as predict developmental outcomes.

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Exposure to Bordetella pertussis adenylate cyclase toxin affects integrin‐mediated adhesion and mechanics in alveolar epithelial cells

Abstract: These results suggest that mechanical and adhesion properties of the cells appear as pertinent markers of cytotoxicity of CyaA toxin.

Cited by 9 publications

References 47 publications

Functional and structural consequences of epithelial cell invasion byBordetella pertussisadenylate cyclase toxin

Functional and structural consequences of epithelial cell invasion byBordetella pertussisadenylate cyclase toxin

A High‐Affinity Calmodulin‐Binding Site in the CyaA Toxin Translocation Domain is Essential for Invasion of Eukaryotic Cells

Biophysical and biomechanical properties of neural progenitor cells as indicators of developmental neurotoxicity

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