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Afadin controls p120-catenin–ZO-1 interactions leading to endothelial barrier enhancement by oxidized phospholipids

Authors

  • Anna A. Birukova,

    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
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  • Panfeng Fu,

    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
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  • Tinghuai Wu,

    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
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  • Oleksii Dubrovskyi,

    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
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  • Nicolene Sarich,

    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
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  • Valery Poroyko,

    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
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  • Konstantin G. Birukov

    Corresponding author
    1. Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, Chicago, Illinois
    • Section of Pulmonary and Critical Medicine, Lung Injury Center, Department of Medicine, University of Chicago, 5841 S. Maryland Ave, Office N-611, Chicago, IL 60637.
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Abstract

Afadin is a novel regulator of epithelial cell junctions assembly. However, its role in the formation of endothelial cell junctions and the regulation of vascular permeability remains obscure. We previously described protective effects of oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (OxPAPC) in the in vitro and in vivo models of lung endothelial barrier dysfunction and acute lung injury, which were mediated by Rac GTPase. This study examined a role of afadin in the OxPAPC-induced enhancement of interactions between adherens junctions and tight junctions as a novel mechanism of endothelial cell (EC) barrier preservation. OxPAPC induced Rap1-dependent afadin accumulation at the cell periphery and Rap1-dependent afadin interaction with adherens junction and tight junction proteins p120-catenin and ZO-1, respectively. Afadin knockdown using siRNA or ectopic expression of afadin mutant lacking Rap1 GTPase binding domain suppressed OxPAPC-induced EC barrier enhancement and abolished barrier protective effects of OxPAPC against thrombin-induced EC permeability. Afadin knockdown also abolished protective effects of OxPAPC against ventilator-induced lung injury in vivo. These results demonstrate for the first time a critical role of afadin in the regulation of vascular barrier function in vitro and in vivo via coordination of adherens junction–tight junction interactions. J. Cell. Physiol. 227: 1883–1890, 2012. © 2011 Wiley Periodicals, Inc.

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