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PMID: 15549136 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Enterotoxigenic Escherichia coli vesicles target toxin delivery into mammalian cells.

The EMBO journal ·Vol. 23 ·No. 23 ·2004-11-24 ·Pages 4538-49

Kesty NC, Mason KM, Reedy M, Miller SE, Kuehn MJ

Abstract

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 degrees C and detected intracellularly at 37 degrees 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.

MeSH Terms
Animals Bacterial Toxins/metabolism Caveolin 1 Caveolins/metabolism Cell Line Cell Membrane/metabolism Endocytosis Enterotoxins/metabolism Escherichia coli/pathogenicity,physiology Escherichia coli Proteins/metabolism Fluorescent Antibody Technique, Indirect HT29 Cells Humans Intestinal Mucosa/cytology,metabolism,ultrastructure Membrane Microdomains/metabolism Mice Microscopy, Electron, Transmission Periplasm/metabolism Transport Vesicles/physiology,ultrastructure Virulence Virulence Factors/metabolism
Chemicals
Bacterial Toxins CAV1 protein, human Cav1 protein, mouse Caveolin 1 Caveolins Enterotoxins Escherichia coli Proteins Virulence Factors heat-labile enterotoxin, E coli
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kesty Nicole C
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
Mason Kevin M
Reedy Mary
Miller Sara E
Kuehn Meta J
References (61)
61 references, click to expand
  1. Escherichia coli cytotoxins and enterotoxins.
    Can J Microbiol. 1992 Jul;38(7):734-46 PMID: 1393838
  2. Meningococcal endotoxin in lethal septic shock plasma studied by gas chromatography, mass-spectrometry, ultracentrifugation, and electron microscopy.
    J Clin Invest. 1992 Mar;89(3):816-23 PMID: 1541674
  3. Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin.
    Microbiol Rev. 1992 Dec;56(4):622-47 PMID: 1480112
  4. Specific adherence of Borrelia burgdorferi extracellular vesicles to human endothelial cells in culture.
    Infect Immun. 1993 Sep;61(9):3892-900 PMID: 8359911
  5. Ultrastructural localization of gangliosides; GM1 is concentrated in caveolae.
    J Histochem Cytochem. 1994 Feb;42(2):155-66 PMID: 8288861
  6. Regulated internalization of caveolae.
    J Cell Biol. 1994 Dec;127(5):1199-215 PMID: 7962085
  7. Protein crystallography and infectious diseases.
    Protein Sci. 1994 Oct;3(10):1670-86 PMID: 7849584
  8. Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion.
    J Bacteriol. 1995 Jul;177(14):3998-4008 PMID: 7608073
  9. The release of outer membrane vesicles from the strains of enterotoxigenic Escherichia coli.
    Microbiol Immunol. 1995;39(7):451-6 PMID: 8569529
  10. Site-directed mutagenic alteration of potential active-site residues of the A subunit of Escherichia coli heat-labile enterotoxin. Evidence for a catalytic role for glutamic acid 112.
    J Biol Chem. 1995 Dec 22;270(51):30545-50 PMID: 8530486
  11. Role of GTP hydrolysis in fission of caveolae directly from plasma membranes.
    Science. 1996 Oct 11;274(5285):239-42 PMID: 8824187
  12. Colonization factors of human enterotoxigenic Escherichia coli (ETEC).
    Trends Microbiol. 1996 Nov;4(11):444-52 PMID: 8950814
  13. Natural release of virulence factors in membrane vesicles by Pseudomonas aeruginosa and the effect of aminoglycoside antibiotics on their release.
    J Antimicrob Chemother. 1997 Nov;40(5):615-21 PMID: 9421308
  14. The World Health Report 1997--conquering suffering, enriching humanity.
    World Health Forum. 1997;18(3-4):248-60 PMID: 9478137
  15. Filipin-dependent inhibition of cholera toxin: evidence for toxin internalization and activation through caveolae-like domains.
    J Cell Biol. 1998 May 18;141(4):905-15 PMID: 9585410
  16. Ganglioside structure dictates signal transduction by cholera toxin and association with caveolae-like membrane domains in polarized epithelia.
    J Cell Biol. 1998 May 18;141(4):917-27 PMID: 9585411
  17. Delivery of the non-membrane-permeative antibiotic gentamicin into mammalian cells by using Shigella flexneri membrane vesicles.
    Antimicrob Agents Chemother. 1998 Jun;42(6):1476-83 PMID: 9624497
  18. Export of virulence genes and Shiga toxin by membrane vesicles of Escherichia coli O157:H7.
    Appl Environ Microbiol. 1999 May;65(5):1843-8 PMID: 10223967
  19. Membrane traffic and the cellular uptake of cholera toxin.
    Biochim Biophys Acta. 1999 Jul 8;1450(3):177-90 PMID: 10395933
  20. Release of Helicobacter pylori vacuolating cytotoxin by both a specific secretion pathway and budding of outer membrane vesicles. Uptake of released toxin and vesicles by gastric epithelium.
    J Pathol. 1999 Jun;188(2):220-6 PMID: 10398168
  21. Structures of gram-negative cell walls and their derived membrane vesicles.
    J Bacteriol. 1999 Aug;181(16):4725-33 PMID: 10438737
  22. Prevalence of toxin types and colonization factors in enterotoxigenic Escherichia coli isolated during a 2-year period from diarrheal patients in Bangladesh.
    J Clin Microbiol. 2000 Jan;38(1):27-31 PMID: 10618058
  23. A role for the bacterial outer membrane in the pathogenesis of Helicobacter pylori infection.
    FEMS Microbiol Lett. 2000 Jan 15;182(2):259-64 PMID: 10620676
  24. Release of gram-negative outer-membrane proteins into human serum and septic rat blood and their interactions with immunoglobulin in antiserum to Escherichia coli J5.
    J Infect Dis. 2000 Mar;181(3):1034-43 PMID: 10720528
  25. Enterotoxigenic Escherichia coli secretes active heat-labile enterotoxin via outer membrane vesicles.
    J Biol Chem. 2000 Apr 28;275(17):12489-96 PMID: 10777535
  26. Lipoproteins from Borrelia burgdorferi applied in liposomes and presented by dendritic cells induce CD8(+) T-lymphocytes in vitro.
    Cell Immunol. 2000 May 1;201(2):124-31 PMID: 10831321
  27. Heterogeneity of detergent-insoluble membranes from human intestine containing caveolin-1 and ganglioside G(M1).
    Am J Physiol Gastrointest Liver Physiol. 2000 Jun;278(6):G895-904 PMID: 10859219
  28. Involvement of cellular caveolae in bacterial entry into mast cells.
    Science. 2000 Aug 4;289(5480):785-8 PMID: 10926542
  29. Caveolin-1 levels are down-regulated in human colon tumors, and ectopic expression of caveolin-1 in colon carcinoma cell lines reduces cell tumorigenicity.
    Cancer Res. 2000 Oct 15;60(20):5870-8 PMID: 11059785
  30. Expression of the endogenous type II secretion pathway in Escherichia coli leads to chitinase secretion.
    EMBO J. 2000 Dec 15;19(24):6697-703 PMID: 11118204
  31. Characteristics of Helicobacter pylori attachment to human primary antral epithelial cells.
    Microbes Infect. 2000 Nov;2(14):1669-76 PMID: 11137040
  32. Iron influences the expression of Helicobacter pylori outer membrane vesicle-associated virulence factors.
    Eur J Gastroenterol Hepatol. 2000 Dec;12(12):1267-73 PMID: 11192314
  33. Microbial pathogenesis: lipid rafts as pathogen portals.
    Curr Biol. 2000 Nov 16;10(22):R823-5 PMID: 11102822
  34. Raft membrane domains: from a liquid-ordered membrane phase to a site of pathogen attack.
    Semin Immunol. 2001 Apr;13(2):89-97 PMID: 11308292
  35. Rapid cycling of lipid raft markers between the cell surface and Golgi complex.
    J Cell Biol. 2001 Apr 30;153(3):529-41 PMID: 11331304
  36. Caveolar endocytosis of simian virus 40 reveals a new two-step vesicular-transport pathway to the ER.
    Nat Cell Biol. 2001 May;3(5):473-83 PMID: 11331875
  37. Caveolae as portals of entry for microbes.
    Microbes Infect. 2001 Jul;3(9):755-61 PMID: 11489424
  38. Endocytosis without clathrin coats.
    Trends Cell Biol. 2001 Oct;11(10):406-12 PMID: 11567873
  39. Outer membrane-like vesicles secreted by Actinobacillus actinomycetemcomitans are enriched in leukotoxin.
    Microb Pathog. 2002 Jan;32(1):1-13 PMID: 11782116
  40. Cell-surface attachment of pedestal-forming enteropathogenic E. coli induces a clustering of raft components and a recruitment of annexin 2.
    J Cell Sci. 2002 Jan 1;115(Pt 1):91-8 PMID: 11801727
  41. Identification of a protein secretory pathway for the secretion of heat-labile enterotoxin by an enterotoxigenic strain of Escherichia coli.
    Proc Natl Acad Sci U S A. 2002 May 14;99(10):7066-71 PMID: 12011463
  42. Plasma membrane cholesterol modulates cellular vacuolation induced by the Helicobacter pylori vacuolating cytotoxin.
    Infect Immun. 2002 Aug;70(8):4112-23 PMID: 12117919
  43. Identification of Escherichia coli outer membrane protein A receptor on human brain microvascular endothelial cells.
    Infect Immun. 2002 Aug;70(8):4556-63 PMID: 12117968
  44. Bacterial surface association of heat-labile enterotoxin through lipopolysaccharide after secretion via the general secretory pathway.
    J Biol Chem. 2002 Sep 6;277(36):32538-45 PMID: 12087095
  45. Association of Helicobacter pylori vacuolating toxin (VacA) with lipid rafts.
    J Biol Chem. 2002 Sep 13;277(37):34642-50 PMID: 12121984
  46. GM1-containing lipid rafts are depleted within clathrin-coated pits.
    Curr Biol. 2003 Apr 15;13(8):686-90 PMID: 12699627
  47. An association of 27- and 40-kDa molecules with glycolipids that bind A-B bacterial enterotoxins to cultured cells.
    Biochim Biophys Acta. 2003 Jun 10;1612(2):186-94 PMID: 12787937
  48. Deep-apical tubules: dynamic lipid-raft microdomains in the brush-border region of enterocytes.
    Biochem J. 2003 Jul 1;373(Pt 1):125-32 PMID: 12689332
  49. Vesicle-mediated export and assembly of pore-forming oligomers of the enterobacterial ClyA cytotoxin.
    Cell. 2003 Oct 3;115(1):25-35 PMID: 14532000
  50. Lipid rafts in epithelial brush borders: atypical membrane microdomains with specialized functions.
    Biochim Biophys Acta. 2003 Oct 31;1617(1-2):1-9 PMID: 14637014
  51. Helicobacter pylori toxin VacA is transferred to host cells via a novel contact-dependent mechanism.
    Cell Microbiol. 2004 Feb;6(2):167-74 PMID: 14706102
  52. Incorporation of heterologous outer membrane and periplasmic proteins into Escherichia coli outer membrane vesicles.
    J Biol Chem. 2004 Jan 16;279(3):2069-76 PMID: 14578354
  53. Lipopolysaccharide 3-deoxy-D-manno-octulosonic acid (Kdo) core determines bacterial association of secreted toxins.
    J Biol Chem. 2004 Feb 27;279(9):8070-5 PMID: 14660669
  54. Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue culture.
    Science. 1974 Jan 25;183(4122):334-6 PMID: 4587905
  55. Amino acid sequence homology between cholera toxin and Escherichia coli heat-labile toxin.
    Nature. 1980 Dec 4;288(5790):499-501 PMID: 7003397
  56. Some characteristics of the outer membrane material released by growing enterotoxigenic Escherichia coli.
    Infect Immun. 1980 Aug;29(2):704-13 PMID: 7011982
  57. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent.
    J Infect Dis. 1987 Mar;155(3):377-89 PMID: 3543152
  58. Comparison of the carbohydrate-binding specificities of cholera toxin and Escherichia coli heat-labile enterotoxins LTh-I, LT-IIa, and LT-IIb.
    Infect Immun. 1988 Jul;56(7):1748-53 PMID: 3290106
  59. Diarrheal disease during Operation Desert Shield.
    N Engl J Med. 1991 Nov 14;325(20):1423-8 PMID: 1656260
  60. Potocytosis: sequestration and transport of small molecules by caveolae.
    Science. 1992 Jan 24;255(5043):410-1 PMID: 1310359
  61. Virulence determinants of Escherichia coli: present knowledge and questions.
    Can J Microbiol. 1992 Jul;38(7):747-52 PMID: 1393839
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600471
Published
2004-11-24
Epub
2004-00-18
Pages
4538-49
Language
English
Region
England
NLM ID
8208664
PMCID
PMC533055
Subset
IM
Grants
NIGMS NIH HHS · R01 GM066140 · United States
NIAID NIH HHS · R03 AI054606 · United States
External Links
PubMed source DOI: 10.1038/sj.emboj.7600471 PMC Full Text View journal details
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