p120 regulates endothelial permeability independently of its NH2 terminus and Rho binding
| Autor: | Crystal R. Herron, Albert B. Reynolds, Anthony M. Lowery, Patricia R. Hollister, Peter A. Vincent |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
rho GTP-Binding Proteins
Delta Catenin animal structures Endothelial permeability Endothelium Physiology Vascular Biology and Microcirculation Plakoglobin Fluorescent Antibody Technique Biology Permeability Adherens junction RNA interference Physiology (medical) medicine Humans Cells Cultured Analysis of Variance Cadherin Catenins Cadherins Molecular biology Cell biology medicine.anatomical_structure Permeability (electromagnetism) Catenin embryonic structures RNA Interference Endothelium Vascular gamma Catenin Cardiology and Cardiovascular Medicine |
| Zdroj: | American journal of physiology. Heart and circulatory physiology. 300(1) |
| ISSN: | 1522-1539 |
| Popis: | The association of p120-catenin (p120) with the juxtamembrane domain (JMD) of vascular endothelial (VE)-cadherin is required to maintain VE-cadherin levels and transendothelial resistance (TEER) of endothelial cell monolayers. To distinguish whether decreased TEER was due to a loss of p120 and not to the decrease in VE-cadherin, we established a system in which p120 was depleted by short hairpin RNA delivered by lentivirus and VE-cadherin was restored via expression of VE-cadherin fused to green fluorescent protein (GFP). Loss of p120 resulted in decreased TEER, which was associated with decreased expression of VE-cadherin, β-catenin, plakoglobin, and α-catenin. Decreased TEER was rescued by restoration of p120 but not by the expression of VE-cadherin-GFP, despite localization of VE-cadherin-GFP at cell-cell borders. Expression of VE-cadherin-GFP restored levels of β-catenin and α-catenin but not plakoglobin, indicating that p120 may be important for recruitment of plakoglobin to the VE-cadherin complex. To evaluate the role of p120 interaction with Rho GTPase in regulating endothelial permeability, we expressed a recombinant form of p120, lacking the NH2terminus and containing alanine substitutions, that eliminates binding of Rho to p120. Expression of this isoform restored expression of the adherens junction complex and rescued permeability as measured by TEER. These results demonstrate that p120 is required for maintaining VE-cadherin expression and TEER independently of its NH2terminus and its role in regulating Rho. |
| Databáze: | OpenAIRE |
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