title: Mechanism of internalization of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans.
abstract: Cytolethal distending toxin (CDT), which is encoded by three genes, cdtA, cdtB and cdtC, is now recognized to have a growing list of biological actions, including inhibition of cell cycle progression, promotion of apoptosis and stimulation of cytokine secretion. It appears that internalization of CDT is essential, at least for cell cycle blockade. Using purified recombinant CDT proteins from the periodontopathic bacterium Actinobacillus actinomycetemcomitans, the authors investigated which combination of toxin proteins produce cell cycle inhibition and which bound and/or entered into host cells. No evidence was found that CdtB bound to HEp-2 human epithelial cells. In contrast, both CdtA and CdtC bound to these cells. Induction of cell cycle arrest required that cells be exposed to both CdtB and CdtC. Pre-exposure of cells to CdtC for as little as 10 min, followed by removal of the free CdtC and addition of exogenous CdtB, resulted in the inhibition of cell cycle progression, suggesting that CdtB could bind to cell-surface-located CdtC. Using various methods to follow internalization of the CDT proteins it was concluded that CdtC acts to bind CdtB at the cell surface and transports it into the cell as a complex via an endosomal pathway blockable by monensin and brefeldin A.
authors.affiliation: Division of Microbial Diseases, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London WC1X 8LD, UK.
authors.forename: Sumio
authors.initials: S
authors.lastname: Akifusa
authors.affiliation: Division of Microbial Diseases, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London WC1X 8LD, UK.
authors.forename: Wendy
authors.initials: W
authors.lastname: Heywood
authors.affiliation: Division of Microbial Diseases, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London WC1X 8LD, UK.
authors.forename: Sean P
authors.initials: SP
authors.lastname: Nair
authors.affiliation: Bone and Mineral Centre, University College London, 5 University Street, London WC1E 6JJ, UK.
authors.forename: Gudrun
authors.initials: G
authors.lastname: Stenbeck
authors.affiliation: Division of Microbial Diseases, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London WC1X 8LD, UK.
authors.forename: Brian
authors.initials: B
authors.lastname: Henderson
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D020126:Brefeldin A / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D006801:Humans; D008985:Monensin / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; D011994:Recombinant Proteins; C067044:cytolethal distending toxin; C447141:cytolethal distending toxin, Actinobacillus actinomycetemcomitans; D020126:Brefeldin A; D008985:Monensin