Bordetella adenylate cyclase toxin promotes calcium entry into both CD11b+ and CD11b- cells through cAMP-dependent L-type-like calcium channels.
Adenylate cyclase toxin (ACT), a 200 kDa protein, is an essential virulence factor for Bordetella pertussis, the bacterium that causes whooping cough. ACT is a member of the pore-forming RTX (repeats-in-toxin) family of proteins that share a characteristic calcium-binding motif of Gly- and Asp-rich nonapeptide repeats and a marked cytolytic or cytotoxic activity. In addition, ACT exhibits a distinctive feature: it has an N-terminal calmodulin-dependent adenylate cyclase domain. Translocation of this domain into the host cytoplasm results in uncontrolled production of cAMP, and it has classically been assumed that this surge in cAMP is the basis for the toxin-mediated killing. Several members of the RTX family of toxins, including ACT, have been shown to induce intracellular calcium increases, through different mechanisms. We show here that ACT stimulates a raft-mediated calcium influx, through its cAMP production activity, that activates PKA, which in turn activates calcium channels with L-type properties. This process is shown to occur both in CD11b(+) and CD11b(-) cells, suggesting a common mechanism, independent of the toxin receptor. We also show that this ACT-induced calcium influx does not correlate with the toxin-induced cytotoxicity.
Martín
César
C
Unidad de Biofísica, Departamento de Bioquímica, Universidad del País Vasco, Centro Mixto CSIC-UPV/EHU, 48080 Bilbao, Spain.
Gómez-Bilbao
Geraxane
G
Ostolaza
Helena
H
D037361:Adenylate Cyclase Toxin / Q000494:pharmacology*; D000818:Animals; D039481:CD11b Antigen / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D002784:Cholesterol / Q000172:deficiency; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D034741:RNA, Small Interfering / Q000378:metabolism; D013696:Temperature
D037361:Adenylate Cyclase Toxin; D039481:CD11b Antigen; D020746:Calcium Channels, L-Type; D034741:RNA, Small Interfering; D002784:Cholesterol; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D002118:Calcium