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Papers In Press, published online ahead of print January 23, 2003
Physiology Dept., University of Tennessee Health Sciences Center, Memphis, TN 38163
Corresponding Author: rkrao{at}physio1.utmem.edu
The activity of Src kinases appears to play a role in both assembly and disassembly of tight junction. However, the role of a specific isoform of Src kinase in regulation of tight junction is not known. In the present study the role of c-Src in regulation of epithelial tight junction was investigated in Caco-2 cell monolayers. Oxidative stress (xanthine oxidase + xanthine) induced an activation and membrane translocation of c-Src. The oxidative stress-induced decrease in transepithelial electrical resistance, increase in inulin permeability, and redistribution of occludin and ZO-1 from the intercellular junctions were prevented by PP2. The rates of oxidative stress-induced activation of c-Src, tyrosine phosphorylation of ZO-1 and b-catenin, decrease in resistance, increase in permeability to inulin, and redistribution of occludin and ZO-1 were significantly greater in cells transfected with wild type c-Src, while it was low in cells transfected with kinase inactive c-SrcK297R mutant, when compared to those in empty vector-transfected cells. The rates of recovery of resistance, increase in barrier to inulin, and reorganization of occludin and ZO-1 into the intercellular junctions during the calcium-induced reassembly of tight junction were much greater in Caco-2 cells transfected with c-SrcK297R as compared to those in cells transfected with empty vector or wild type c-Src. These results show that the dominant-negative expression of kinase inactive c-Src delays the oxidative stress-induced disruption of tight junction, and accelerates calcium-induced assembly of tight junction in Caco-2 cells, and demonstrate that oxidative stress-induced disruption of tight junction is mediated by the activation of c-Src.
J. Biol. Chem, 10.1074/jbc.M211710200
Submitted on November 18, 2002
Revised on January 21, 2003
Accepted on January 22, 2003
Expression of kinase-inactive c-Src delays oxidative stress-induced disassembly and accelerates calcium-mediated reassembly of tight junctions in Caco-2 cell monolayer
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