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

doi:10.1128/IAI.69.3.1329-1336.2001

This Article

	Full Text
	Full Text (PDF)
	Alert me when this article is cited
	Alert me if a correction is posted

Services

	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new issues of the journal
	Download to citation manager
	Reprints and Permissions
	Copyright Information
	Books from ASM Press
	MicrobeWorld

Citing Articles

	Citing Articles via HighWire
	Citing Articles via Google Scholar

Google Scholar

	Articles by Nusrat, A.
	Articles by Parkos, C. A.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Nusrat, A.
	Articles by Parkos, C. A.

Previous Article | Next Article

Infection and Immunity, March 2001, p. 1329-1336, Vol. 69, No. 3
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.3.1329-1336.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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

A. Nusrat,¹^,^* C. von Eichel-Streiber,² J. R. Turner,³^,⁴ P. Verkade,⁵ J. L. Madara,¹ and C. A. Parkos¹

Epithelial Pathobiology Research Unit, Department of Pathology, Emory University School of Medicine, Atlanta, Georgia¹; Institut fur Medizinische Mikrobiologie und Hygiene, Universität Mainz, Mainz,² Cell Biology and Biophysics Program, EMBL, Heidelberg,³ and Max Planck Institute of Molecular Cell Biology and Genetics, Dresden,⁴ Germany; and Department of Pathology, Wayne State University, Detroit, Michigan⁵

Received 28 July 2000/Returned for modification 14 September 2000/Accepted 20 November 2000

The anaerobic bacterium Clostridium difficile is the etiologic agent of pseudomembranous colitis. C. difficile toxins TcdAand TcdB are UDP-glucosyltransferases that monoglucosylate andthereby inactivate the Rho family of GTPases (W. P. Ciesla, Jr.,and D. A. Bobak, J. Biol. Chem. 273:16021-16026, 1998). We utilizedpurified reference toxins of C. difficile, TcdA-10463 (TcdA) andTcdB-10463 (TcdB), and a model intestinal epithelial cell lineto characterize their influence on tight-junction (TJ) organizationand hence to analyze the mechanisms by which they contribute tothe enhanced paracellular permeability and disease pathophysiologyof pseudomembranous colitis. The increase in paracellular permeabilityinduced by TcdA and TcdB was associated with disorganization ofapical and basal F-actin. F-actin restructuring was paralleledby dissociation of occludin, ZO-1, and ZO-2 from the lateral TJmembrane without influencing the subjacent adherens junction protein,E-cadherin. In addition, we observed decreased association ofactin with the TJ cytoplasmic plaque protein ZO-1. Differentialdetergent extraction and fractionation in sucrose density gradientsrevealed TcdB-induced redistribution of occludin and ZO-1 fromdetergent-insoluble fractions constituting "raft-like" membranemicrodomains, suggesting an important role of Rho proteins inmaintaining the association of TJ proteins with such microdomains.These toxin-mediated effects on actin and TJ structure providea mechanism for early events in the pathophysiology of pseudomembranouscolitis.

^* Corresponding author. Mailing address: Department of Pathology and Laboratory Medicine, Emory University, WMRB 2335, 1639Pierce Dr., Atlanta, GA 30322. Phone: (404) 727-8543. Fax: (404)727-3321. E-mail: anusrat{at}emory.edu.

Infection and Immunity, March 2001, p. 1329-1336, Vol. 69, No. 3
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.3.1329-1336.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

Casselli, T., Lynch, T., Southward, C. M., Jones, B. W., DeVinney, R. (2008). Vibrio parahaemolyticus Inhibition of Rho Family GTPase Activation Requires a Functional Chromosome I Type III Secretion System. Infect. Immun. 76: 2202-2211 [Abstract] [Full Text]
Moser, L. A., Carter, M., Schultz-Cherry, S. (2007). Astrovirus Increases Epithelial Barrier Permeability Independently of Viral Replication. J. Virol. 81: 11937-11945 [Abstract] [Full Text]
Beau, I., Cotte-Laffitte, J., Amsellem, R., Servin, A. L. (2007). A Protein Kinase A-Dependent Mechanism by Which Rotavirus Affects the Distribution and mRNA Level of the Functional Tight Junction-Associated Protein, Occludin, in Human Differentiated Intestinal Caco-2 Cells. J. Virol. 81: 8579-8586 [Abstract] [Full Text]
Chen, M. L., Ge, Z., Fox, J. G., Schauer, D. B. (2006). Disruption of Tight Junctions and Induction of Proinflammatory Cytokine Responses in Colonic Epithelial Cells by Campylobacter jejuni. Infect. Immun. 74: 6581-6589 [Abstract] [Full Text]
Shen, L., Black, E. D., Witkowski, E. D., Lencer, W. I., Guerriero, V., Schneeberger, E. E., Turner, J. R. (2006). Myosin light chain phosphorylation regulates barrier function by remodeling tight junction structure. J. Cell Sci. 119: 2095-2106 [Abstract] [Full Text]
Nguyen, H.-V., Chen, J.-L., Zhong, J., Kim, K.-J., Crandall, E. D., Borok, Z., Chen, Y., Ann, D. K. (2006). SUMOylation Attenuates Sensitivity toward Hypoxia- or Desferroxamine-Induced Injury by Modulating Adaptive Responses in Salivary Epithelial Cells. Am. J. Pathol. 168: 1452-1463 [Abstract] [Full Text]
Klingberg, T. D., Pedersen, M. H., Cencic, A., Budde, B. B. (2005). Application of Measurements of Transepithelial Electrical Resistance of Intestinal Epithelial Cell Monolayers To Evaluate Probiotic Activity. Appl. Environ. Microbiol. 71: 7528-7530 [Abstract] [Full Text]
Pechine, S., Janoir, C., Collignon, A. (2005). Variability of Clostridium difficile Surface Proteins and Specific Serum Antibody Response in Patients with Clostridium difficile-Associated Disease. J. Clin. Microbiol. 43: 5018-5025 [Abstract] [Full Text]
Bruewer, M., Utech, M., Ivanov, A. I., Hopkins, A. M., Parkos, C. A., Nusrat, A. (2005). Interferon-{gamma} induces internalization of epithelial tight junction proteins via a macropinocytosis-like process. FASEB J. 19: 923-933 [Abstract] [Full Text]
Guo, X., Rao, J. N., Liu, L., Zou, T., Keledjian, K. M., Boneva, D., Marasa, B. S., Wang, J.-Y. (2005). Polyamines are necessary for synthesis and stability of occludin protein in intestinal epithelial cells. Am. J. Physiol. Gastrointest. Liver Physiol. 288: G1159-G1169 [Abstract] [Full Text]
Voth, D. E., Ballard, J. D. (2005). Clostridium difficile Toxins: Mechanism of Action and Role in Disease. Clin. Microbiol. Rev. 18: 247-263 [Abstract] [Full Text]
Lynch, T., Livingstone, S., Buenaventura, E., Lutter, E., Fedwick, J., Buret, A. G., Graham, D., DeVinney, R. (2005). Vibrio parahaemolyticus Disruption of Epithelial Cell Tight Junctions Occurs Independently of Toxin Production. Infect. Immun. 73: 1275-1283 [Abstract] [Full Text]
Solomon, K., Webb, J., Ali, N., Robins, R. A., Mahida, Y. R. (2005). Monocytes Are Highly Sensitive to Clostridium difficile Toxin A-Induced Apoptotic and Nonapoptotic Cell Death. Infect. Immun. 73: 1625-1634 [Abstract] [Full Text]
Go, M., Kojima, T., Takano, K.-i., Murata, M., Ichimiya, S., Tsubota, H., Himi, T., Sawada, N. (2004). Expression and Function of Tight Junctions in the Crypt Epithelium of Human Palatine Tonsils. J. Histochem. Cytochem. 52: 1627-1638 [Abstract] [Full Text]
Li, N., Lewis, P., Samuelson, D., Liboni, K., Neu, J. (2004). Glutamine regulates Caco-2 cell tight junction proteins. Am. J. Physiol. Gastrointest. Liver Physiol. 287: G726-G733 [Abstract] [Full Text]
Bruewer, M., Hopkins, A. M., Hobert, M. E., Nusrat, A., Madara, J. L. (2004). RhoA, Rac1, and Cdc42 exert distinct effects on epithelial barrier via selective structural and biochemical modulation of junctional proteins and F-actin. Am. J. Physiol. Cell Physiol. 287: C327-C335 [Abstract] [Full Text]
Ivanov, A. I., McCall, I. C., Parkos, C. A., Nusrat, A. (2004). Role for Actin Filament Turnover and a Myosin II Motor in Cytoskeleton-driven Disassembly of the Epithelial Apical Junctional Complex. Mol. Biol. Cell 15: 2639-2651 [Abstract] [Full Text]
Ivanov, A. I., Nusrat, A., Parkos, C. A. (2004). Endocytosis of Epithelial Apical Junctional Proteins by a Clathrin-mediated Pathway into a Unique Storage Compartment. Mol. Biol. Cell 15: 176-188 [Abstract] [Full Text]
Bruewer, M., Luegering, A., Kucharzik, T., Parkos, C. A., Madara, J. L., Hopkins, A. M., Nusrat, A. (2003). Proinflammatory Cytokines Disrupt Epithelial Barrier Function by Apoptosis-Independent Mechanisms. J. Immunol. 171: 6164-6172 [Abstract] [Full Text]
Kansagra, K., Stoll, B., Rognerud, C., Niinikoski, H., Ou, C.-N., Harvey, R., Burrin, D. (2003). Total parenteral nutrition adversely affects gut barrier function in neonatal piglets. Am. J. Physiol. Gastrointest. Liver Physiol. 285: G1162-G1170 [Abstract] [Full Text]
Chin, A. C., Vergnolle, N., MacNaughton, W. K., Wallace, J. L., Hollenberg, M. D., Buret, A. G. (2003). Proteinase-activated receptor 1 activation induces epithelial apoptosis and increases intestinal permeability. Proc. Natl. Acad. Sci. USA 100: 11104-11109 [Abstract] [Full Text]
Yoo, J., Nichols, A., Mammen, J., Calvo, I., Song, J. C., Worrell, R. T., Matlin, K., Matthews, J. B. (2003). Bryostatin-1 enhances barrier function in T84 epithelia through PKC-dependent regulation of tight junction proteins. Am. J. Physiol. Cell Physiol. 285: C300-C309 [Abstract] [Full Text]
Spyres, L. M., Daniel, J., Hensley, A., Qa'Dan, M., Ortiz-Leduc, W., Ballard, J. D. (2003). Mutational Analysis of the Enzymatic Domain of Clostridium difficile Toxin B Reveals Novel Inhibitors of the Wild-Type Toxin. Infect. Immun. 71: 3294-3301 [Abstract] [Full Text]
Liu, T. S., Musch, M. W., Sugi, K., Walsh-Reitz, M. M., Ropeleski, M. J., Hendrickson, B. A., Pothoulakis, C., Lamont, J. T., Chang, E. B. (2003). Protective role of HSP72 against Clostridium difficile toxin A-induced intestinal epithelial cell dysfunction. Am. J. Physiol. Cell Physiol. 284: C1073-C1082 [Abstract] [Full Text]
Rubenstein, N. M., Guan, Y., Woo, P. L., Firestone, G. L. (2003). Glucocorticoid Down-regulation of RhoA Is Required for the Steroid-induced Organization of the Junctional Complex and Tight Junction Formation in Rat Mammary Epithelial Tumor Cells. J. Biol. Chem. 278: 10353-10360 [Abstract] [Full Text]
Berkes, J, Viswanathan, V K, Savkovic, S D, Hecht, G (2003). Intestinal epithelial responses to enteric pathogens: effects on the tight junction barrier, ion transport, and inflammation. Gut 52: 439-451 [Abstract] [Full Text]
Hopkins, A. M., Walsh, S. V., Verkade, P., Boquet, P., Nusrat, A. (2003). Constitutive activation of Rho proteins by CNF-1 influences tight junction structure and epithelial barrier function. J. Cell Sci. 116: 725-742 [Abstract] [Full Text]
Tafazoli, F., Magnusson, K.-E., Zheng, L. (2003). Disruption of Epithelial Barrier Integrity by Salmonella enterica Serovar Typhimurium Requires Geranylgeranylated Proteins. Infect. Immun. 71: 872-881 [Abstract] [Full Text]
Edens, H. A., Levi, B. P., Jaye, D. L., Walsh, S., Reaves, T. A., Turner, J. R., Nusrat, A., Parkos, C. A. (2002). Neutrophil Transepithelial Migration: Evidence for Sequential, Contact-Dependent Signaling Events and Enhanced Paracellular Permeability Independent of Transjunctional Migration. J. Immunol. 169: 476-486 [Abstract] [Full Text]
Chen, M. L., Pothoulakis, C., LaMont, J. T. (2002). Protein Kinase C Signaling Regulates ZO-1 Translocation and Increased Paracellular Flux of T84 Colonocytes Exposed to Clostridium difficile Toxin A. J. Biol. Chem. 277: 4247-4254 [Abstract] [Full Text]
Abreu, M. T., Arnold, E. T., Chow, J. Y. C., Barrett, K. E. (2001). Phosphatidylinositol 3-Kinase-dependent Pathways Oppose Fas-induced Apoptosis and Limit Chloride Secretion in Human Intestinal Epithelial Cells. IMPLICATIONS FOR INFLAMMATORY DIARRHEAL STATES. J. Biol. Chem. 276: 47563-47574 [Abstract] [Full Text]

J. Bacteriol.	J. Virol.	Eukaryot. Cell

Microbiol. Mol. Biol. Rev.	Clin. Vaccine Immunol.	All ASM Journals