Journal Article

Clostridium difficile Toxin CDT Induces Formation of Microtubule-Based Protrusions and Increases Adherence of Bacteria

Stecher, Bärbel, Schwan, Carsten, Rohde, Manfred, van Ham, Marco, Aktories, Klaus, Tzivelekidis, Tina, Hardt, Wolf-Dietrich, Wehland, Jürgen

2009

Overview

Clostridium difficile causes antibiotic-associated diarrhea and pseudomembranous colitis by production of the Rho GTPase-glucosylating toxins A and B. Recently emerging hypervirulent Clostridium difficile strains additionally produce the binary ADP-ribosyltransferase toxin CDT (Clostridium difficile transferase), which ADP-ribosylates actin and inhibits actin polymerization. Thus far, the role of CDT as a virulence factor is not understood. Here we report by using time-lapse- and immunofluorescence microscopy that CDT and other binary actin-ADP-ribosylating toxins, including Clostridium botulinum C2 toxin and Clostridium perfringens iota toxin, induce redistribution of microtubules and formation of long (up to >150 microm) microtubule-based protrusions at the surface of intestinal epithelial cells. The toxins increase the length of decoration of microtubule plus-ends by EB1/3, CLIP-170 and CLIP-115 proteins and cause redistribution of the capture proteins CLASP2 and ACF7 from microtubules at the cell cortex into the cell interior. The CDT-induced microtubule protrusions form a dense meshwork at the cell surface, which wrap and embed bacterial cells, thereby largely increasing the adherence of Clostridia. The study describes a novel type of microtubule structure caused by less efficient microtubule capture and offers a new perspective for the pathogenetic role of CDT and other binary actin-ADP-ribosylating toxins in host-pathogen interactions.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Actin Cytoskeleton - drug effects / Actin Cytoskeleton - metabolism / ADP Ribose Transferases - metabolism / ADP Ribose Transferases - pharmacology / ADP Ribose Transferases - physiology / Animals / Bacteria / Bacterial Adhesion - drug effects / Bacterial Adhesion - physiology / Bacterial toxins / Bacterial Toxins - metabolism / Bacterial Toxins - pharmacology / Caco-2 Cells / Cell Biology/Cytoskeleton / Cell culture / Cell Surface Extensions - drug effects / Cell Surface Extensions - metabolism / Clostridium botulinum / Clostridium difficile / Clostridium difficile - enzymology / Clostridium difficile - physiology / Clostridium perfringens / Cytoskeleton / Dose-Response Relationship, Drug / Experiments / Gastroenterology and Hepatology/Gastrointestinal Infections / Germ-Free Life / Health aspects / HT29 Cells / Humans / Inflammatory bowel disease / Mice / Microbiology / Microbiology/Cellular Microbiology and Pathogenesis / Microscopy / Microtubules / Microtubules - drug effects / Microtubules - metabolism / Physiological aspects / Proteins / Rats / Rats, Wistar / Toxins