Expression of kinase-inactive c-Src delays oxidative stress-induced disassembly and accelerates calcium-mediated reassembly of tight junctions in the Caco-2 cell monolayer

Shyamali Basuroy, Parimal Sheth, Dhan Kuppuswamy, Sundar Balasubramanian, Ramesh M. Ray, Radhakrishna Rao

Research output: Contribution to journalArticle

119 Citations (Scopus)

Abstract

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 β-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, whereas it was low in cells transfected with kinaseinactive c-SrcK297R mutant, when compared with 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 with 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.

Original languageEnglish (US)
Pages (from-to)11916-11924
Number of pages9
JournalJournal of Biological Chemistry
Volume278
Issue number14
DOIs
StatePublished - Apr 4 2003

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Oxidative stress
Caco-2 Cells
Tight Junctions
Monolayers
Occludin
Oxidative Stress
Phosphotransferases
Inulin
Calcium
src-Family Kinases
Chemical activation
Intercellular Junctions
Catenins
Phosphorylation
Permeability
Xanthine
Acoustic impedance
Xanthine Oxidase
Tyrosine
Protein Isoforms

All Science Journal Classification (ASJC) codes

  • Biochemistry

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Expression of kinase-inactive c-Src delays oxidative stress-induced disassembly and accelerates calcium-mediated reassembly of tight junctions in the Caco-2 cell monolayer. / Basuroy, Shyamali; Sheth, Parimal; Kuppuswamy, Dhan; Balasubramanian, Sundar; Ray, Ramesh M.; Rao, Radhakrishna.

In: Journal of Biological Chemistry, Vol. 278, No. 14, 04.04.2003, p. 11916-11924.

Research output: Contribution to journalArticle

Basuroy, Shyamali ; Sheth, Parimal ; Kuppuswamy, Dhan ; Balasubramanian, Sundar ; Ray, Ramesh M. ; Rao, Radhakrishna. / Expression of kinase-inactive c-Src delays oxidative stress-induced disassembly and accelerates calcium-mediated reassembly of tight junctions in the Caco-2 cell monolayer. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 14. pp. 11916-11924.
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