tailieunhanh - Báo cáo khoa hoc : Cellular vacuolation induced by Clostridium perfringens epsilon-toxin

The epsilon-toxin ofClostridium perfringensforms a heptamer in the mem-branes of Madin–Darby canine kidney cells, leading to cell death. Here, we report that it caused the vacuolation of Madin–Darby canine kidney cells. The toxin induced vacuolation in a dose-dependent and time-dependent manner. | IFEBS Journal Cellular vacuolation induced by Clostridium perfringens epsilon-toxin It D KI I 1xzi lal I 1 I I I I I I 2kAI II 1 1 1 X z I I- I . 1 Masahiro Nagahama Yukari Itohayashi Hideki Hara2 Masahiro Higashihara Yusuke Fukatani T QI I ihics T aLafiichi1 l loQO f olzo o1 1 zt Ĩ lx zx rtltax achi1 Inhi m M al a ia A a3 and Ilin c ZJ x I I t zx 11 Teiuhisa Takagishi Iviasaiaka Oda Keiko Kobayashi IChiio Nakagawa and Jun sakurai 1 Department of Microbiology Faculty of PharmaceuticalSciences Tokushima Bunri University Japan 2 Department of Microbiology Kyoto University Graduate Schoolof Medicine Japan 3 Section of BacterialPathogenesis Graduate Schoolof Medicaland DentalSciences Tokyo Medicaland DentalUniversity Japan Keywords C. perfringens epsilon-toxin endocytosis endosomes lysosomes vacuolation Correspondence M. Nagahama Department of Microbiology Faculty of Pharmaceutical Sciences Tokushima Bunri University Yamashiro-cho Tokushima 770-8514 Japan Fax 81 088 655 3051 Tel 81 088 622 9611 E-mail nagahama@ Received 8 March 2011 revised 27 June 2011 accepted 20 July 2011 doi The epsilon-toxin of Clostridium perfringens forms a heptamer in the membranes of Madin-Darby canine kidney cells leading to cell death. Here we report that it caused the vacuolation of Madin-Darby canine kidney cells. The toxin induced vacuolation in a dose-dependent and time-dependent manner. The monomer of the toxin formed oligomers on lipid rafts in membranes of the cells. Methyl-b-cyclodextrin and poly ethylene glycol 4000 inhibited the vacuolation. Epsilon-toxin was internalized into the cells. Confocal microscopy revealed that the internalized toxin was transported from early endosomes early endosome antigen 1 staining to late endosomes and lysosomes lysosomal-associated membrane protein 2 staining and then distributed to the membranes of vacuoles. Furthermore the vacuolation was inhibited by bafilomycin A1 a V-type ATPase inhibitor and .