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VEGF controls endothelial-cell permeability by promoting the β-arrestin-dependent endocytosis of VE-cadherin

Nature Cell Biology volume 8, pages 1223–1234 (2006)Cite this article

Abstract

How vascular endothelial growth factor (VEGF) induces vascular permeability, its first described function, remains poorly understood. Here, we provide evidence of a novel signalling pathway by which VEGF stimulation promotes the rapid endocytosis of a key endothelial cell adhesion molecule, VE-cadherin, thereby disrupting the endothelial barrier function. This process is initiated by the activation of the small GTPase Rac by VEGFR-2 through the Src-dependent phosphorylation of Vav2, a guanine nucleotide-exchange factor. Rac activation, in turn, promotes the p21-activated kinase (PAK)-mediated phosphorylation of a highly conserved motif within the intracellular tail of VE-cadherin. Surprisingly, this results in the recruitment of β-arrestin2 to serine-phosphorylated VE-cadherin, thereby promoting its internalization into clathrin-coated vesicles and the consequent disassembly of intercellular junctions. Ultimately, this novel biochemical route by which VEGF promotes endothelial permeability through the β-arrestin2-dependent endocytosis of VE-cadherin may help identify new therapeutic targets for the treatment of many human diseases that are characterized by vascular leakage.

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Figure 1: VEGF promotes VE-cadherin endocytosis.
Figure 2: VEGF induces the endocytosis of VE-cadherin by phosphorylating Vav2 through Src.
Figure 3: A highly conserved SVR motif in human VE-cadherin is a potential target for PAK phosphorylation.
Figure 4: Ser 665 regulates VE-cadherin endocytosis and VEGF-induced permeability.
Figure 5: A role for Ser 665 in VEGF induced coclustering of β-arrestin2 with VE-cadherin.
Figure 6: β-arrestin2 is involved in VEGF-induced VE-cadherin endocytosis and permeability.
Figure 7: Schematic representation of a model of the molecular mechanisms by which VEGF promotes VE-cadherin internalization and vascular permeability by VE-cadherin receptor β-arrestin-dependent endocytosis (RADE) on the activation of the Vav2–Rac–PAK signalling pathway through VEGFR-2 and Src.

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Acknowledgements

We are grateful: to W.A. Muller (Weill Medical College of Cornell University, New York, NY) for the human VE-cadherin cDNA; to J.L. Benovic (Department of Biochemistry and Molecular Biology Thomas Jefferson University, Philadelphia, PA) for the arrestin antibodies; to M.C. Frame (The Beatson Institute for Cancer Research, Glasgow, UK) for the Src-GFP plasmid; and to L. Lamalice and J. Huot (Université de Laval, Québec, Canada) for the VEGFR-2–HA plasmid. We also thank D. Martin for helpful advice on shRNA vectors, R. Castilho for genomic expression data, C. Murga and S. Fukuhara for preparation of GFP–β-arrestin2 plasmid. We also thank J. Basile and T. Bugge for critical reading of the manuscript. J.G. is supported by a fellowship from Fondation pour la Recherche Médicale (http://www.frm.org). This research was partially supported by the Intramural Research Program of the National Institutes of Health (NIH), National Institute of Dental and Craniofacial research (NIDCR).

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  1. Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, DHHS, Bethesda, 20892–4340, MD, USA

    Julie Gavard & J. Silvio Gutkind

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J.G. and J.S.G. planned the experimental design, analysed data and wrote the paper. J.G. conducted the experiments.

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Correspondence to J. Silvio Gutkind.

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Gavard, J., Gutkind, J. VEGF controls endothelial-cell permeability by promoting the β-arrestin-dependent endocytosis of VE-cadherin. Nat Cell Biol 8, 1223–1234 (2006). https://doi.org/10.1038/ncb1486

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