title: Perturbation of mitochondrial structure and function plays a central role in Actinobacillus actinomycetemcomitans leukotoxin-induced apoptosis.
abstract: Certain pore-forming bacterial toxins, including the leukotoxin (Ltx) produced by Actinobacillus actinomycetemcomitans, induce apoptosis in susceptible target cells. Although binding to the target cell surface represents the first step in the initiation of this process, the downstream events leading to toxin-induced apoptotic cell death have not been identified. Perturbation of mitochondrial function has been shown to have a major role in regulating progression of apoptosis initiated by exposure to numerous stimuli. Using Ltx as a model, the aim of this study was to evaluate whether induction of apoptosis by pore-forming toxins follows a similar paradigm. After exposure to Ltx, Epstein-Barr virus transformed B cells (JY cell line) exhibited the classical morphological features of apoptosis including decreased cell size, plasma membrane blebbing, selective alterations in plasma membrane permeability and condensation of nuclear DNA. The morphologic changes were accompanied by swelling of the mitochondria, a decrease in mitochondrial transmembrane potential (Psi(m)), hyperproduction of reactive oxygen intermediates (ROIs) and release of cytochrome c from the intermembrane space. Subsequently, we detected activation of the c ysteine asp artate-specific prote ases (caspases)-3 and -9, cleavage of the nuclear DNA repair enzyme, poly(ADP-ribose)polymerase (PARP) and internucleosomal DNA fragmentation. These results indicate that perturbation of mitochondrial structure and function, in concert with activation of specific caspases, initiate the effector phase of Ltx-induced apoptosis.
authors.affiliation: Department of Periodontics, Leon Levy Research Center for Oral Biology, University of Pennsylvania, 4010 Locust Street, Philadelphia, PA 19104-6002, USA. jkorosto@pobox.upenn.edu
authors.forename: J
authors.initials: J
authors.lastname: Korostoff
authors.forename: N
authors.initials: N
authors.lastname: Yamaguchi
authors.forename: M
authors.initials: M
authors.lastname: Miller
authors.forename: I
authors.initials: I
authors.lastname: Kieba
authors.forename: E T
authors.initials: ET
authors.lastname: Lally
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans / Q000187:drug effects* / Q000502:physiology; D017209:Apoptosis; D020169:Caspases / Q000032:analysis; D002460:Cell Line; D003574:Cytochrome c Group / Q000032:analysis; D003600:Cytosol / Q000201:enzymology; D003603:Cytotoxins / Q000494:pharmacology*; D053938:DNA Fragmentation; D004789:Enzyme Activation; D005098:Exotoxins / Q000494:pharmacology*; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects; D008564:Membrane Potentials / Q000187:drug effects; D008854:Microscopy, Electron; D008928:Mitochondria / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D010539:Permeability / Q000187:drug effects; D011065:Poly(ADP-ribose) Polymerases / Q000032:analysis / Q000187:drug effects
chemical_list: D003574:Cytochrome c Group; D003603:Cytotoxins; D005098:Exotoxins; C018579:leukotoxin; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases