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A more recent version of this article appeared on October 1, 2005
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Submitted on March 8, 2005
Revised on June 28, 2005
Accepted on July 19, 2005
induced Endocytosis of Tight Junction Proteins: Myosin II-dependent Vacuolarization of the Apical Plasma Membrane
*Epithelial Pathobiology Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322; Department of General Surgery, University of Muenster, 48149 Muenster, Germany; Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 142290 Pushchino, Russia; Department of Pathology, The University of Chicago, Chicago, IL 60637; ||Welsh School of Pharmacy, University of Wales, Cardiff CF10 3XF, Wales, United Kingdom
Monitoring Editor: Keith Mostov
Disruption of epithelial barrier by proinflammatory cytokines such as IFN-
represents a major pathophysiological consequence of intestinal inflammation. We have previously shown that IFN- increases paracellular permeability in model T84 epithelial cells by inducing endocytosis of tight junction (TJ) proteins occludin, JAM-A and claudin-1. The present study was designed to dissect mechanisms of IFN--induced endocytosis of epithelial TJ proteins. IFN- treatment of T84 cells resulted in internalization of TJ proteins into large actin-coated vacuoles which originated from the apical plasma membrane and resembled the vacuolar apical compartment (VAC) previously observed in epithelial cells that lose cell polarity. The IFN- dependent formation of VACs required ATPase activity of a myosin II motor but was not dependent on rapid turnover of F-actin. In addition, activated myosin II was observed to colocalize with VACs after IFN- exposure. Pharmacological analyses revealed that formation of VACs and endocytosis of TJ proteins was mediated by Rho-associated kinase (ROCK) but not myosin light chain kinase (MLCK). Furthermore, IFN- treatment resulted in activation of Rho GTPase and induced expressional up-regulation of ROCK. These results, for the first time, suggest that IFN- induces endocytosis of epithelial TJ proteins via RhoA/ROCK-mediated, myosin II-dependent formation of VACs.
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