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| Subject Categories: Membranes & Transport | Microbiology & Pathogens |
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| The EMBO Journal
(2004) 23, 4538–4549, doi:10.1038/sj.emboj.7600471 Published online 18 November 2004 |
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| Enterotoxigenic Escherichia coli vesicles target toxin delivery into mammalian cells |
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| Nicole C Kesty1, Kevin M Mason1, 4, Mary Reedy2, Sara E Miller3 and Meta J Kuehn1 |
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1 Department of Biochemistry, Duke University Medical Center, Durham, NC, USA 2 Department of Cell Biology, Duke University Medical Center, Durham, NC, USA 3 Department of Pathology, Duke University Medical Center, Durham, NC, USA To whom correspondence should be addressed Meta J Kuehn, Department of Biochemistry, Duke University Medical Center, 132 Nanaline Duke, Box 3711, Durham, NC 27710, USA. Tel.: +1 919 684 2545; Fax: +1 919 684 8885; E-mail: meta.kuehn@duke.edu 4 Present address: Department of Pediatrics, Columbus Children's Research Institute, The Ohio State University College of Medicine and Public Health, Columbus, OH 43205, USA Received 18 February 2004; Accepted 12 October 2004; Published online 18 November 2004. |
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| Abstract |
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| Enterotoxigenic Escherichia coli (ETEC) is a prevalent cause of traveler's diarrhea and infant mortality in third-world countries. Heat-labile enterotoxin (LT) is secreted from ETEC via vesicles composed of outer membrane and periplasm. We investigated the role of ETEC vesicles in pathogenesis by analyzing vesicle association and entry into eukaryotic cells. Fluorescently labeled vesicles from LT-producing and LT-nonproducing strains were compared in their ability to bind adrenal and intestinal epithelial cells. ETEC-derived vesicles, but not control nonpathogen-derived vesicles, associated with cells in a time-, temperature-, and receptor-dependent manner. Vesicles were visualized on the cell surface at 4°C and detected intracellularly at 37°C. ETEC vesicle endocytosis depended on cholesterol-rich lipid rafts. Entering vesicles partially colocalized with caveolin, and the internalized vesicles accumulated in a nonacidified compartment. We conclude that ETEC vesicles serve as specifically targeted transport vehicles that mediate entry of active enterotoxin and other bacterial envelope components into host cells. These data demonstrate a role in virulence for ETEC vesicles. |
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| Keywords: endocytosis, lipid raft, LT, secretion, toxin |
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