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J. Biol. Chem., Vol. 276, Issue 13, 10423-10431, March 30, 2001
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From the In epithelial and endothelial cells, tight
junctions regulate the paracellular permeability of ions and proteins.
Disruption of tight junctions by inflammation is often associated with
tissue edema, but regulatory mechanisms are not fully understood. Using ECV304 cells as a model system, lysophosphatidic acid and histamine were found to increase the paracellular permeability of the tracer horseradish peroxidase. Cytoskeletal changes induced by these agents
included stimulation of stress fiber formation and myosin light chain
phosphorylation. Additionally, occludin, a tight junction protein, was
a target for signaling events triggered by lysophosphatidic acid and
histamine, events that resulted in its phosphorylation. A
dominant-negative mutant of RhoA, RhoA T19N, or a specific inhibitor of
Rho-activated kinases, Y-27632, prevented stress fiber formation, myosin light chain phosphorylation, occludin phosphorylation, and the
increase in tracer flux in response to lysophosphatidic acid. In
contrast, although RhoA T19N and Y-27632 blocked the cytoskeletal
events induced by histamine, they had no effect on the stimulation of
occludin phosphorylation or increased tracer flux, indicating that
occludin phosphorylation may regulate tight junction permeability
independently of cytoskeletal events. Thus, occludin is a target for
receptor-initiated signaling events regulating its phosphorylation, and
this phosphorylation may be a key regulator of tight junction permeability.
Regulation of Tight Junction Permeability and Occludin
Phosphorylation by RhoA-p160ROCK-dependent and
-independent Mechanisms*
§,,,,
,, and First Department of Internal Medicine, Kobe
University School of Medicine, 7-5-2 Kusunoki-cho, Chuo-ku, Kobe
650-0017, Japan, Department of Cell Biology, Faculty of
Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan, and ¶ Eisai London Research Laboratories Ltd., University
College London, Gower Street, London WC1E 6BT, United Kingdom
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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