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J Biol Chem, Vol. 273, Issue 29, 18122-18129, July 17, 1998

Aerolysin Induces G-protein Activation and Ca²⁺ Release from Intracellular Stores in Human Granulocytes

Karl-Heinz Krause, Marc Fivaz^§, Antoinette Monod, and F. Gisou van der Goot^§

From the  Infectious Diseases Division, University Hospital, 1211 Geneva 14, Switzerland and the ^§ Department of Biochemistry, University of Geneva, 30 quai E. Ansermet, 1211 Geneva 4, Switzerland

Aerolysin is a pore-forming toxin that plays a key role in the pathogenesis of Aeromonas hydrophila infections. In this study, we have analyzed the effect of aerolysin on human granulocytes (HL-60 cells). Proaerolysin could bind to these cells, was processed into active aerolysin, and led to membrane depolarization, indicating that granulocytes are potential targets for this toxin. Fura-2 measurements were used to analyze the effect of aerolysin on cytosolic [Ca²⁺] homeostasis. As expected for a pore-forming toxin, aerolysin addition led to Ca²⁺ influx across the plasma membrane. In addition, the toxin triggered Ca²⁺ release from agonist and thapsigargin-sensitive intracellular Ca²⁺ stores. This Ca²⁺ release was independent of the aerolysin-induced Ca²⁺ influx and occurred in two kinetically distinct phases: an initial rapid and transient phase and a second, more sustained, phase. The first, but not the second phase was sensitive to pertussis toxin. Activation of pertussis toxin-sensitive G-proteins appeared to be a consequence of pore formation, rather than receptor activation through aerolysin-binding, as it: (i) was not observed with a binding competent, insertion-incompetent aerolysin mutant, (ii) had a marked lag time, and (iii) was also observed in response to other bacterial pore-forming toxins (staphylococcal -toxin, streptolysin O) which are thought to bind to different receptors. G-protein activation through pore-forming toxins stimulated cellular functions, as evidenced by pertussis toxin-sensitive chemotaxis. Our results demonstrate that granulocytes are potential target cells for aerolysin and that in these cells, Ca²⁺ signaling in response to a pore-forming toxin involves G-protein-dependent cell activation and Ca²⁺ release from intracellular stores.

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