scholarly journals Bordetella adenylate cyclase toxin is a unique ligand of the integrin complement receptor 3

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Radim Osicka ◽  
Adriana Osickova ◽  
Shakir Hasan ◽  
Ladislav Bumba ◽  
Jiri Cerny ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Whooping Cough ◽  
Complement Receptor ◽  
Ligand Interaction ◽  
Downstream Signaling ◽  
Cell Functions ◽  
Natural Ligand ◽  
Adenylate Cyclase Toxin ◽  
Complement Receptor 3 ◽  
Signaling Receptors

Integrins are heterodimeric cell surface adhesion and signaling receptors that are essential for metazoan existence. Some integrins contain an I-domain that is a major ligand binding site. The ligands preferentially engage the active forms of the integrins and trigger signaling cascades that alter numerous cell functions. Here we found that the adenylate cyclase toxin (CyaA), a key virulence factor of the whooping cough agent Bordetella pertussis, preferentially binds an inactive form of the integrin complement receptor 3 (CR3), using a site outside of its I-domain. CyaA binding did not trigger downstream signaling of CR3 in human monocytes and CyaA-catalyzed elevation of cAMP effectively blocked CR3 signaling initiated by a natural ligand. This unprecedented type of integrin-ligand interaction distinguishes CyaA from all other known ligands of the I-domain-containing integrins and provides a mechanistic insight into the previously observed central role of CyaA in the pathogenesis of B. pertussis.

scholarly journals Author response: Bordetella adenylate cyclase toxin is a unique ligand of the integrin complement receptor 3

2015 ◽  
Author(s):  
Radim Osicka ◽  
Adriana Osickova ◽  
Shakir Hasan ◽  
Ladislav Bumba ◽  
Jiri Cerny ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Author Response ◽  
Complement Receptor ◽  
Adenylate Cyclase Toxin ◽  
Complement Receptor 3

scholarly journals Almost half of the RTX domain is dispensable for complement receptor 3 binding and cell-invasive activity of the adenylate cyclase toxin

2021 ◽  
pp. 100833
Author(s):  
Carlos Angel Espinosa-Vinals ◽  
Jiri Masin ◽  
Jana Holubova ◽  
Ondrej Stanek ◽  
David Jurnecka ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Complement Receptor ◽  
Adenylate Cyclase Toxin ◽  
Complement Receptor 3

scholarly journals Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding

FEBS Letters ◽  
2014 ◽  
Vol 589 (3) ◽  
pp. 374-379 ◽  
Author(s):  
Shakir Hasan ◽  
Adriana Osickova ◽  
Ladislav Bumba ◽  
Petr Novák ◽  
Peter Sebo ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Complement Receptor ◽  
Adenylate Cyclase Toxin ◽  
Complement Receptor 3

scholarly journals Phospholipase A activity of adenylate cyclase toxin mediates translocation of its adenylate cyclase domain

2017 ◽  
Vol 114 (33) ◽  
pp. E6784-E6793 ◽  
Author(s):  
David González-Bullón ◽  
Kepa B. Uribe ◽  
César Martín ◽  
Helena Ostolaza
Keyword(s):  
Cell Membrane ◽  
Adenylate Cyclase ◽  
Molecular Mechanism ◽  
Catalytic Domain ◽  
Whooping Cough ◽  
Phospholipase A ◽  
Host Cell Membrane ◽  
Adenylate Cyclase Toxin ◽  
Causative Bacterium ◽  
Domain Transfer

Adenylate cyclase toxin (ACT or CyaA) plays a crucial role in respiratory tract colonization and virulence of the whooping cough causative bacteriumBordetella pertussis. Secreted as soluble protein, it targets myeloid cells expressing the CD11b/CD18 integrin and on delivery of its N-terminal adenylate cyclase catalytic domain (AC domain) into the cytosol, generates uncontrolled toxic levels of cAMP that ablates bactericidal capacities of phagocytes. Our study deciphers the fundamentals of the heretofore poorly understood molecular mechanism by which the ACT enzyme domain directly crosses the host cell membrane. By combining molecular biology, biochemistry, and biophysics techniques, we discover that ACT has intrinsic phospholipase A (PLA) activity, and that such activity determines AC translocation. Moreover, we show that elimination of the ACT–PLA activity abrogates ACT toxicity in macrophages, particularly at toxin concentrations close to biological reality of bacterial infection. Our data support a molecular mechanism in which in situ generation of nonlamellar lysophospholipids by ACT–PLA activity into the cell membrane would form, likely in combination with membrane-interacting ACT segments, a proteolipidic toroidal pore through which AC domain transfer could directly take place. Regulation of ACT–PLA activity thus emerges as novel target for therapeutic control of the disease.

scholarly journals Quantification of the Adenylate Cyclase Toxin of Bordetella pertussisIn Vitroand during Respiratory Infection

2013 ◽  
Vol 81 (5) ◽  
pp. 1390-1398 ◽  
Author(s):  
Joshua C. Eby ◽  
Mary C. Gray ◽  
Jason M. Warfel ◽  
Christopher D. Paddock ◽  
Tara F. Jones ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Respiratory Tract ◽  
Whooping Cough ◽  
Extensive Study ◽  
Target Cells ◽  
Human Infants ◽  
Content Type ◽  
Adenylate Cyclase Toxin

ABSTRACTWhooping cough results from infection of the respiratory tract withBordetella pertussis, and the secreted adenylate cyclase toxin (ACT) is essential for the bacterium to establish infection. Despite extensive study of the mechanism of ACT cytotoxicity and its effects over a range of concentrationsin vitro, ACT has not been observed or quantifiedin vivo, and thus the concentration of ACT at the site of infection is unknown. The recently developed baboon model of infection mimics the prolonged cough and transmissibility of pertussis, and we hypothesized that measurement of ACT in nasopharyngeal washes (NPW) from baboons, combined with human andin vitrodata, would provide an estimate of the ACT concentration in the airway during infection. NPW contained up to ∼108CFU/mlB. pertussisand 1 to 5 ng/ml ACT at the peak of infection. Nasal aspirate specimens from two human infants with pertussis contained bacterial concentrations similar to those in the baboons, with 12 to 20 ng/ml ACT. When ∼108CFU/ml of a laboratory strain ofB. pertussiswas culturedin vitro, ACT production was detected in 60 min and reached a plateau of ∼60 ng/ml in 6 h. Furthermore, when bacteria were brought into close proximity to target cells by centrifugation, intoxication was increased 4-fold. Collectively, these data suggest that at the bacterium-target cell interface during infection of the respiratory tract, the concentration of ACT can exceed 100 ng/ml, providing a reference point for future studies of ACT and pertussis pathogenesis.

scholarly journals Albumin, in the Presence of Calcium, Elicits a Massive Increase in Extracellular Bordetella Adenylate Cyclase Toxin

2017 ◽  
Vol 85 (6) ◽  
Author(s):  
Laura A. Gonyar ◽  
Mary C. Gray ◽  
Gregory J. Christianson ◽  
Borna Mehrad ◽  
Erik L. Hewlett
Keyword(s):  
Adenylate Cyclase ◽  
Whooping Cough ◽  
Critical Factor ◽  
The United States ◽  
Transcriptional Level ◽  
Content Type ◽  
Novel Host ◽  
Fetal Bovine ◽  
Adenylate Cyclase Toxin ◽  
Myeloid Leukocytes

ABSTRACT Pertussis (whooping cough), caused by Bordetella pertussis, is resurging in the United States and worldwide. Adenylate cyclase toxin (ACT) is a critical factor in establishing infection with B. pertussis and acts by specifically inhibiting the response of myeloid leukocytes to the pathogen. We report here that serum components, as discovered during growth in fetal bovine serum (FBS), elicit a robust increase in the amount of ACT, and ≥90% of this ACT is localized to the supernatant, unlike growth without FBS, in which ≥90% is associated with the bacterium. We have found that albumin, in the presence of physiological concentrations of calcium, acts specifically to enhance the amount of ACT and its localization to the supernatant. Respiratory secretions, which contain albumin, promote an increase in amount and localization of active ACT that is comparable to that elicited by serum and albumin. The response to albumin is not mediated through regulation of ACT at the transcriptional level or activation of the Bvg two-component system. As further illustration of the specificity of this phenomenon, serum collected from mice that lack albumin does not stimulate an increase in ACT. These data, demonstrating that albumin and calcium act synergistically in the host environment to increase production and release of ACT, strongly suggest that this phenomenon reflects a novel host-pathogen interaction that is central to infection with B. pertussis and other Bordetella species.

scholarly journals The role of four cholesterol-recognition motifs localized between amino acid residues 400-550 in regulating translocation and lytic activity of Adenylate Cyclase Toxin

2021 ◽  
Author(s):  
Jone Amuategi ◽  
Rocio Alonso ◽  
Helena Ostolaza
Keyword(s):  
Adenylate Cyclase ◽  
Whooping Cough ◽  
Cell Signalling ◽  
Lipid Vesicles ◽  
Cell Types ◽  
Target Cells ◽  
Amino Acid Residues ◽  
Cyclase Domain ◽  
Adenylate Cyclase Toxin ◽  
Recognition Motifs

Adenylate Cyclase Toxin (ACT or CyaA) is an important virulence factor secreted by Bordetella pertussis, the bacterium causative of whooping cough, playing an essential role in the establishment of infection in the respiratory tract. ACT is a pore-forming cytolysin belonging to the RTX (Repeats in ToXin) family of leukotoxins, capable of permeabilizing several cell types and pure lipid vesicles. Besides, the toxin delivers its N-terminal adenylate cyclase domain into the target cytosol, where catalyzes the conversion of ATP into cAMP, which affects cell signalling. In this study we have made two major observations. First, we show that ACT binds free cholesterol, and identify in its sequence 38 potential cholesterol-recognition motifs. Second, we reveal that four of those motifs are real, functional cholesterol-binding sites. Mutations of the central phenylalanine residues in said motifs have an important impact on the ACT lytic and translocation activities, suggesting their direct intervention in cholesterol recognition and toxin functionality. From our data a likely transmembrane topology can be inferred for the ACT helices constituting the translocation and the hydrophobic regions. From this topology a simple and plausible mechanism emerges by which ACT could translocate its AC domain into target cells, challenging previous views in the field. Blocking the ACT-cholesterol interactions might thus be an effective approach for inhibiting ACT toxicity on cells, and this could help in mitigating the severity of pertussis disease in humans.

scholarly journals Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme–translocating segment of Bordetella adenylate cyclase toxin

2020 ◽  
Vol 295 (28) ◽  
pp. 9349-9365
Author(s):  
Jiri Masin ◽  
Adriana Osickova ◽  
David Jurnecka ◽  
Nela Klimova ◽  
Humaira Khaliq ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Adenylate Cyclase ◽  
Adenylyl Cyclase ◽  
Specific Capacity ◽  
Target Cells ◽  
Jurkat T Cells ◽  
Membrane Interaction ◽  
The Family ◽  
Adenylate Cyclase Toxin ◽  
Complement Receptor 3

The Bordetella adenylate cyclase toxin-hemolysin (CyaA) and the α-hemolysin (HlyA) of Escherichia coli belong to the family of cytolytic pore-forming Repeats in ToXin (RTX) cytotoxins. HlyA preferentially binds the αLβ2 integrin LFA-1 (CD11a/CD18) of leukocytes and can promiscuously bind and also permeabilize many other cells. CyaA bears an N-terminal adenylyl cyclase (AC) domain linked to a pore-forming RTX cytolysin (Hly) moiety, binds the complement receptor 3 (CR3, αMβ2, CD11b/CD18, or Mac-1) of myeloid phagocytes, penetrates their plasma membrane, and delivers the AC enzyme into the cytosol. We constructed a set of CyaA/HlyA chimeras and show that the CyaC-acylated segment and the CR3-binding RTX domain of CyaA can be functionally replaced by the HlyC-acylated segment and the much shorter RTX domain of HlyA. Instead of binding CR3, a CyaA1-710/HlyA411-1024 chimera bound the LFA-1 receptor and effectively delivered AC into Jurkat T cells. At high chimera concentrations (25 nm), the interaction with LFA-1 was not required for CyaA1-710/HlyA411-1024 binding to CHO cells. However, interaction with the LFA-1 receptor strongly enhanced the specific capacity of the bound CyaA1-710/HlyA411-1024 chimera to penetrate cells and deliver the AC enzyme into their cytosol. Hence, interaction of the acylated segment and/or the RTX domain of HlyA with LFA-1 promoted a productive membrane interaction of the chimera. These results help delimit residues 400–710 of CyaA as an “AC translocon” sufficient for translocation of the AC polypeptide across the plasma membrane of target cells.

scholarly journals Bordetella pertussis Inhibition of Interleukin-12 (IL-12) p70 in Human Monocyte-Derived Dendritic Cells Blocks IL-12 p35 through Adenylate Cyclase Toxin-Dependent Cyclic AMP Induction

2006 ◽  
Vol 74 (5) ◽  
pp. 2831-2838 ◽  
Author(s):  
Fabiana Spensieri ◽  
Giorgio Fedele ◽  
Cecilia Fazio ◽  
Maria Nasso ◽  
Paola Stefanelli ◽  
...  
Keyword(s):  
Immune Response ◽  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Bordetella Pertussis ◽  
Interleukin 12 ◽  
Protein Chain ◽  
Cell Functions ◽  
A Subunit ◽  
Adenylate Cyclase Toxin ◽  
The Impact

ABSTRACT Bordetella pertussis, the causative agent of whooping cough, possesses an array of virulence factors, including adenylate cyclase toxin (ACT), relevant in the establishment of infection. Here we better define the impact of cyclic AMP (cAMP) intoxication due to the action of ACT on dendritic cell (DC)-driven immune response, by infecting monocyte-derived DC (MDDC) with an ACT-deficient B. pertussis mutant (ACT−18HS19) or its parental strain (WT18323). Both strains induced MDDC maturation and antigen-presenting cell functions; however, only ACT−18HS19 infected MDDC-induced production of interleukin-12 (IL-12) p70. Gene expression analysis of the IL-12 cytokine family subunits revealed that both strains induced high levels of p40 (protein chain communal to IL-12 p70 and IL-23) as well as p19, a subunit of IL-23. Conversely only ACT−18HS19 infection induced consistent transcription of IL-12 p35, a subunit of IL-12 p70. Addition of the cAMP analogous d-butyril-cAMP (d-cAMP) abolished IL-12 p70 production and IL-12 p35 expression in ACT−18HS19-infected MDDC. ACT−18HS19 infection induced the expression of the transcription factors interferon regulatory factor 1 (IRF-1) and IRF-8 and of beta interferon, involved in IL-12 p35 regulation, and the expression of these genes was inhibited by d-cAMP addition and in WT18323-infected MDDC. The concomitant expression of IL-12 p70 and IL-23 allowed ACT−18HS19 to trigger a more pronounced T helper 1 polarization compared to WT18323. The present study suggests that ACT-dependent cAMP induction leads to the inhibition of pathways ultimately leading to IL-12 p35 production, thus representing a mechanism for B. pertussis to escape the host immune response.

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