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Molecular and Cellular Biology, March 2008, p. 1657-1668, Vol. 28, No. 5
0270-7306/08/$08.00+0     doi:10.1128/MCB.00695-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Contribution of Annexin 2 to the Architecture of Mature Endothelial Adherens Junctions{triangledown} ,{dagger}

Stéphanie Heyraud,1 Michel Jaquinod,2 Claire Durmort,3 Emilie Dambroise,1 Evelyne Concord,1 Jean Patrick Schaal,4 Philippe Huber,1 and Danielle Gulino-Debrac1*

Laboratoire de Physiopathologie Vasculaire, INSERM U882,1 EDyP, INSERM UM201,2 LIM, IBS, CNRS UMR 5075, CEA, Direction of Life Sciences, and Joseph Fourier University, 38054 Grenoble, France,3 Département de Gynécologie Obstétrique, Centre Hospitalier Universitaire Michallon, BP 217, 38043 Grenoble Cedex 09, France4

Received 20 April 2007/ Returned for modification 29 June 2007/ Accepted 11 December 2007

The vascular endothelial cadherin (VE-cad)-based complex is involved in the maintenance of vascular endothelium integrity. Using immunoprecipitation experiments, we have demonstrated that, in confluent human umbilical vein endothelial cells, the VE-cad-based complex interacts with annexin 2 and that annexin 2 translocates from the cytoplasm to the cell-cell contact sites as cell confluence is established. Annexin 2, located in cholesterol rafts, binds to both the actin cytoskeleton and the VE-cad-based complex so the complex is docked to cholesterol rafts. These multiple connections prevent the lateral diffusion of the VE-cad-based complex, thus strengthening adherens junctions in the ultimate steps of maturation. Moreover, we observed that the down-regulation of annexin 2 by small interfering RNA induces a delocalization of VE-cad from adherens junctions and consequently a destabilization of these junctions. Furthermore, our data indicate that the decoupling of the annexin 2/p11 complex from the VE-cad-based junction, triggered by vascular endothelial growth factor treatment, facilitates the switch from a quiescent to an immature state.

* Corresponding author. Mailing address: iRTSV, APV, INSERM U882, CEA-Grenoble, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France. Phone: 33 4 38 78 24 92. Fax: 33 4 38 78 49 64. E-mail: danielle.gulino{at}cea.fr

{triangledown} Published ahead of print on 26 December 2007.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, March 2008, p. 1657-1668, Vol. 28, No. 5
0270-7306/08/$08.00+0     doi:10.1128/MCB.00695-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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