The oxidative-stress-induced alteration in paracellular junctional complexes was analysed in Caco-2 cell monolayer. Oxidative stress induced a rapid increase in tyrosine phosphorylation of occludin, zonula occludens (ZO)-1, E-cadherin and β-catenin. An oxidative-stress-induced decrease in transepithelial electrical resistance was associated with a redistribution of occludin—ZO-1 and E-cadherin—β-catenin complexes from the intercellular junctions. Genistein, a tyrosine kinase inhibitor, prevented the oxidative-stress-induced decrease in resistance and redistribution of protein complexes. Occludin, ZO-1, E-cadherin and β-catenin in the Triton-insoluble cytoskeletal fraction were reduced by oxidative stress, which was prevented by genistein. Oxidative stress also reduced the co-immunoprecipitation of ZO-1 with occludin, which was prevented by genistein. Co-immunoprecipitation of β-catenin with E-cadherin was unaffected by oxidative stress or genistein. ZO-1, E-cadherin and β-catenin in the plasma membrane or membrane-cytoskeleton were either slightly reduced or unaffected by oxidative stress or genistein. These results show that oxidative stress induces tyrosine phosphorylation and cellular redistribution of occludin—ZO-1 and E-cadherin—β-catenin complexes by a tyrosine-kinase-dependent mechanism.
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December 2002
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Research Article|
December 01 2002
Tyrosine phosphorylation and dissociation of occludin–ZO-1 and E-cadherin–β-catenin complexes from the cytoskeleton by oxidative stress
Radhakrishna K. RAO;
Radhakrishna K. RAO
1
∗Department of Physiology, University of Tennessee Health Sciences Center, 894 Union Avenue, Memphis, TN 38163, U.S.A.
1To whom correspondence should be addressed (e-mail rkrao@physio1.utmem.edu).
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Shyamali BASUROY;
Shyamali BASUROY
∗Department of Physiology, University of Tennessee Health Sciences Center, 894 Union Avenue, Memphis, TN 38163, U.S.A.
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Vijay U. RAO;
Vijay U. RAO
†Department of Pediatrics, Medical University of South Carolina, 171 Ashley Avenue, Charleston, SC, U.S.A.,
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Karl J. KARNAKY, Jr;
Karl J. KARNAKY, Jr
†Department of Pediatrics, Medical University of South Carolina, 171 Ashley Avenue, Charleston, SC, U.S.A.,
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Akshay GUPTA
Akshay GUPTA
‡Department of Cell Biology, Medical University of South Carolina, 171 Ashley Avenue, Charleston, SC, U.S.A.
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Publisher: Portland Press Ltd
Received:
December 08 2001
Revision Received:
July 03 2002
Accepted:
August 08 2002
Accepted Manuscript online:
August 08 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 368 (2): 471–481.
Article history
Received:
December 08 2001
Revision Received:
July 03 2002
Accepted:
August 08 2002
Accepted Manuscript online:
August 08 2002
Citation
Radhakrishna K. RAO, Shyamali BASUROY, Vijay U. RAO, Karl J. KARNAKY, Akshay GUPTA; Tyrosine phosphorylation and dissociation of occludin–ZO-1 and E-cadherin–β-catenin complexes from the cytoskeleton by oxidative stress. Biochem J 1 December 2002; 368 (2): 471–481. doi: https://doi.org/10.1042/bj20011804
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