Clostridium Difficile Toxins Disrupt Epithelial Barrier Function by Altering Membrane Microdomain Localization of Tight Junction Proteins

Overview

Journal Infect Immun

Publisher American Society for Microbiology

Specialty Allergy & Immunology

Date 2001 Feb 17

PMID 11179295

Citations 131

Authors

A Nusrat

C von Eichel-Streiber

J R Turner

P Verkade

J L Madara

C A Parkos

Affiliations

Soon will be listed here.

Abstract

The anaerobic bacterium Clostridium difficile is the etiologic agent of pseudomembranous colitis. C. difficile toxins TcdA and TcdB are UDP-glucosyltransferases that monoglucosylate and thereby inactivate the Rho family of GTPases (W. P. Ciesla, Jr., and D. A. Bobak, J. Biol. Chem. 273:16021-16026, 1998). We utilized purified reference toxins of C. difficile, TcdA-10463 (TcdA) and TcdB-10463 (TcdB), and a model intestinal epithelial cell line to characterize their influence on tight-junction (TJ) organization and hence to analyze the mechanisms by which they contribute to the enhanced paracellular permeability and disease pathophysiology of pseudomembranous colitis. The increase in paracellular permeability induced by TcdA and TcdB was associated with disorganization of apical and basal F-actin. F-actin restructuring was paralleled by dissociation of occludin, ZO-1, and ZO-2 from the lateral TJ membrane without influencing the subjacent adherens junction protein, E-cadherin. In addition, we observed decreased association of actin with the TJ cytoplasmic plaque protein ZO-1. Differential detergent extraction and fractionation in sucrose density gradients revealed TcdB-induced redistribution of occludin and ZO-1 from detergent-insoluble fractions constituting "raft-like" membrane microdomains, suggesting an important role of Rho proteins in maintaining the association of TJ proteins with such microdomains. These toxin-mediated effects on actin and TJ structure provide a mechanism for early events in the pathophysiology of pseudomembranous colitis.

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References

Nusrat A, Parkos C, Verkade P, Foley C, Liang T, Eastburn K . Tight junctions are membrane microdomains. J Cell Sci. 2000; 113 ( Pt 10):1771-81. DOI: 10.1242/jcs.113.10.1771. View

Wong V . Phosphorylation of occludin correlates with occludin localization and function at the tight junction. Am J Physiol. 1998; 273(6):C1859-67. DOI: 10.1152/ajpcell.1997.273.6.C1859. View

Madara J, Stafford J, Dharmsathaphorn K, Carlson S . Structural analysis of a human intestinal epithelial cell line. Gastroenterology. 1987; 92(5 Pt 1):1133-45. DOI: 10.1016/s0016-5085(87)91069-9. View

Madara J . Intestinal absorptive cell tight junctions are linked to cytoskeleton. Am J Physiol. 1987; 253(1 Pt 1):C171-5. DOI: 10.1152/ajpcell.1987.253.1.C171. View

Gumbiner B . Structure, biochemistry, and assembly of epithelial tight junctions. Am J Physiol. 1987; 253(6 Pt 1):C749-58. DOI: 10.1152/ajpcell.1987.253.6.C749. View

Madara J, MOORE R, Carlson S . Alteration of intestinal tight junction structure and permeability by cytoskeletal contraction. Am J Physiol. 1987; 253(6 Pt 1):C854-61. DOI: 10.1152/ajpcell.1987.253.6.C854. View

Madara J, Stafford J, Barenberg D, Carlson S . Functional coupling of tight junctions and microfilaments in T84 monolayers. Am J Physiol. 1988; 254(3 Pt 1):G416-23. DOI: 10.1152/ajpgi.1988.254.3.G416. View

Madara J . Tight junction dynamics: is paracellular transport regulated?. Cell. 1988; 53(4):497-8. DOI: 10.1016/0092-8674(88)90562-4. View

Hecht G, Pothoulakis C, Lamont J, Madara J . Clostridium difficile toxin A perturbs cytoskeletal structure and tight junction permeability of cultured human intestinal epithelial monolayers. J Clin Invest. 1988; 82(5):1516-24. PMC: 442717. DOI: 10.1172/JCI113760. View

10.

STEVENSON B, Anderson J, Bullivant S . The epithelial tight junction: structure, function and preliminary biochemical characterization. Mol Cell Biochem. 1988; 83(2):129-45. DOI: 10.1007/BF00226141. View

11.

von Eichel-Streiber C, Harperath U, Bosse D, HADDING U . Purification of two high molecular weight toxins of Clostridium difficile which are antigenically related. Microb Pathog. 1987; 2(5):307-18. DOI: 10.1016/0882-4010(87)90073-8. View

12.

Shapiro M, Matthews J, Hecht G, Delp C, Madara J . Stabilization of F-actin prevents cAMP-elicited Cl- secretion in T84 cells. J Clin Invest. 1991; 87(6):1903-9. PMC: 296941. DOI: 10.1172/JCI115215. View

13.

Parkos C, Delp C, Arnaout M, Madara J . Neutrophil migration across a cultured intestinal epithelium. Dependence on a CD11b/CD18-mediated event and enhanced efficiency in physiological direction. J Clin Invest. 1991; 88(5):1605-12. PMC: 295682. DOI: 10.1172/JCI115473. View

14.

Hecht G, Koutsouris A, Pothoulakis C, Lamont J, Madara J . Clostridium difficile toxin B disrupts the barrier function of T84 monolayers. Gastroenterology. 1992; 102(2):416-23. DOI: 10.1016/0016-5085(92)90085-d. View

15.

Brown D, Rose J . Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell. 1992; 68(3):533-44. DOI: 10.1016/0092-8674(92)90189-j. View

16.

Parkos C, Colgan S, Delp C, Arnaout M, Madara J . Neutrophil migration across a cultured epithelial monolayer elicits a biphasic resistance response representing sequential effects on transcellular and paracellular pathways. J Cell Biol. 1992; 117(4):757-64. PMC: 2289458. DOI: 10.1083/jcb.117.4.757. View

17.

Schneeberger E, LYNCH R . Structure, function, and regulation of cellular tight junctions. Am J Physiol. 1992; 262(6 Pt 1):L647-61. DOI: 10.1152/ajplung.1992.262.6.L647. View

18.

Kaoutzani P, Parkos C, Madara J . Isolation of plasma membrane fractions from the intestinal epithelial model T84. Am J Physiol. 1993; 264(5 Pt 1):C1327-35. DOI: 10.1152/ajpcell.1993.264.5.C1327. View

19.

Sargiacomo M, Sudol M, Tang Z, Lisanti M . Signal transducing molecules and glycosyl-phosphatidylinositol-linked proteins form a caveolin-rich insoluble complex in MDCK cells. J Cell Biol. 1993; 122(4):789-807. PMC: 2119592. DOI: 10.1083/jcb.122.4.789. View

20.

Anderson R . Caveolae: where incoming and outgoing messengers meet. Proc Natl Acad Sci U S A. 1993; 90(23):10909-13. PMC: 47891. DOI: 10.1073/pnas.90.23.10909. View