Oligomerization is involved in pore formation by Bordetella adenylate cyclase toxin

Vojtova-Vodolanova, Jana and Basler, Marek and Osicka, Radim and Knapp, Oliver and Maier, Elke and Cerny, Jan and Benada, Oldrich and Benz, Roland and Sebo, Peter. (2009) Oligomerization is involved in pore formation by Bordetella adenylate cyclase toxin. FASEB Journal, 23 (9). pp. 2831-2843.

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Official URL: http://edoc.unibas.ch/49542/

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The Bordetella adenylate cyclase-hemolysin (CyaA, ACT, or AC-Hly) is a multifunctional toxin. Simultaneously with promoting calcium ion entry, CyaA delivers into host cells an adenylate cyclase enzyme (AC) and permeabilizes cell membrane by forming small cation-selective pores. Indirect evidence suggested that these two activities were accomplished by different membrane-inserted CyaA conformers, one acting as an AC-delivering monomer and the other as an uncharacterized pore-forming oligomer. We tested this model by directly detecting toxin oligomers in cell membrane and by assessing oligomerization of specific mutants with altered pore-forming properties. CyaA oligomers were revealed in sheep erythrocyte membranes by immunogold labeling and directly demonstrated by pulldown of membrane-inserted CyaA together with biotinylated CyaA-AC(-) toxoid. Membrane oligomers of CyaA could also be resolved by nondenaturing electrophoresis of mild detergent extracts of erythrocytes. Furthermore, CyaA mutants exhibiting enhanced (E581K) or reduced (E570K+E581P) specific hemolytic and pore-forming activity were found to exhibit also a correspondingly enhanced or reduced propensity to form oligomers in erythrocyte membranes. On the other hand, processed CyaA, with the AC domain cleaved off by erythrocyte proteases, was detected only in a monomeric form excluded from the oligomers of unprocessed CyaA. These results provide the first direct evidence that oligomerization is involved in formation of CyaA pores in target membranes and that translocation of the AC domain across cell membrane may be accomplished by monomeric CyaA.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Infection Biology > Infection Biology (Basler)
UniBasel Contributors:Basler, Marek
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Federation of American Society of Experimental Biology
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:29 Nov 2017 08:23
Deposited On:29 Nov 2017 08:23

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