Membrane Permeabilization by Bordetella Adenylate Cyclase Toxin Involves Pores of Tunable Size.

Abstract

RTX (Repeats in ToXin) pore-forming toxins constitute an expanding family of exoproteins secreted by many Gram-negative bacteria and involved in infectious diseases caused by said pathogens. Despite the relevance in the host/pathogen interactions, the structure and characteristics of the lesions formed by these toxins remain enigmatic. Here, we capture the first direct nanoscale pictures of lytic pores formed by an RTX toxin, the Adenylate cyclase (ACT), secreted by the whooping cough bacterium Bordetella pertussis. We reveal that ACT associates into growing-size oligomers of variable stoichiometry and heterogeneous architecture (lines, arcs, and rings) that pierce the membrane, and that, depending on the incubation time and the toxin concentration, evolve into large enough "holes" so as to allow the flux of large molecular mass solutes, while vesicle integrity is preserved. We also resolve ACT assemblies of similar variable stoichiometry in the cell membrane of permeabilized target macrophages, proving that our model system recapitulates the process of ACT permeabilization in natural membranes. Based on our data we propose a non-concerted monomer insertion and sequential mechanism of toroidal pore formation by ACT. A size-tunable pore adds a new regulatory element to ACT-mediated cytotoxicity, with different pore sizes being putatively involved in different physiological scenarios or cell types.

• Authors: David González-Bullón, Kepa B Uribe, Eneko Largo, Garazi Guembelzu, Aritz B García-Arribas, César Martín, Helena Ostolaza
• Journal: Biomolecules (Biomolecules) Vol. 9 Issue 5 (05 10 2019) ISSN: 2218-273X [Electronic] Switzerland
• PMID: 31083482 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Adenylate Cyclase Toxin
• Pore Forming Cytotoxic Proteins

Topics

• Adenylate Cyclase Toxin (chemistry, metabolism, toxicity)
• Animals
• Bordetella pertussis (enzymology, pathogenicity)
• Cell Line
• Cell Membrane (metabolism)
• Cell Membrane Permeability
• Macrophages (microbiology)
• Mice
• Pore Forming Cytotoxic Proteins (chemistry, metabolism, toxicity)
• Protein Binding
• Protein Multimerization