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First published online 20 July 2004
doi: 10.1242/jcs.01246

Journal of Cell Science 117, 3911-3922 (2004)
Published by The Company of Biologists 2004
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Research Article

Shiga toxin binding to globotriaosyl ceramide induces intracellular signals that mediate cytoskeleton remodeling in human renal carcinoma-derived cells

Hisami Takenouchi1,2, Nobutaka Kiyokawa1,*, Tomoko Taguchi1, Jun Matsui1, Yohko U. Katagiri1, Hajime Okita1, Kenji Okuda2 and Junichiro Fujimoto1

1 Department of Developmental Biology, National Research Institute for Child Health and Development, 3-35-31, Taishido, Setagaya-ku, Tokyo, 154-8567, Japan
2 Department of Bacteriology, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan

* Author for correspondence (e-mail: nkiyokawa{at}nch.go.jp)

Accepted 6 April 2004

Shiga toxin is a bacterial toxin consisting of A and B subunits. Generally, the essential cytotoxicity of the toxin is thought to be mediated by the A subunit, which possesses RNA cleavage activity and thus induces protein synthesis inhibition. We previously reported, however, that the binding of the Shiga toxin 1-B subunit to globotriaosyl ceramide, a functional receptor for Shiga toxin, induces intracellular signals in a manner that is dependent on glycolipid-enriched membrane domains, or lipid rafts. Although the precise role of this signaling mechanism is not known, here we report that Shiga-toxin-mediated intracellular signals induce cytoskeleton remodeling in ACHN cells derived from renal tubular epithelial carcinoma. Using confocal laser scanning microscopy, we observed that Shiga toxin 1-B treatment induces morphological changes in ACHN cells in a time-dependent manner. In addition, the morphological changes were accompanied by the redistribution of a number of proteins, including actin, ezrin, CD44, vimentin, cytokeratin, paxillin, FAK, and {alpha}- and {gamma}-tubulins, all of which are involved in cytoskeletal organization. The transient phosphorylation of ezrin and paxillin was also observed during the course of protein redistribution. Experiments using inhibitors for a variety of kinases suggested the involvement of lipid rafts, Src family protein kinase, PI 3-kinase, and RHO-associated kinase in Shiga toxin 1-B-induced ezrin phosphorylation. Shiga toxin 1-B-induced cytoskeletal remodeling should provide an in vitro model that can be used to increase our understanding of the pathogenesis of Shiga-toxin-mediated cell injury and the role of lipid-raft-mediated cell signaling in cytoskeletal remodeling.

Key words: Shiga toxin, Globotriaosylceramide, Cytoskeleton


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