HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 34, 30376-30383, August 26, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/34/30376    most recent M502844200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Andreeva, A. V. Articles by Voyno-Yasenetskaya, T. Search for Related Content PubMed PubMed Citation Articles by Andreeva, A. V. Articles by Voyno-Yasenetskaya, T. Social Bookmarking                      What's this?

G₁₂ Interaction with SNAP Induces VE-cadherin Localization at Endothelial Junctions and Regulates Barrier Function^*

Alexandra V. Andreeva, Mikhail A. Kutuzov, Rita Vaiskunaite, Jasmina Profirovic, Thomas E. Meigs¶, Sanda Predescu, Asrar B. Malik, and Tatyana Voyno-Yasenetskaya, An established investigator of the American Heart Association||

From the Department of Pharmacology, College of Medicine, University of Illinois, Chicago, Illinois 60612 and ^¶University of North Carolina, Asheville, North Carolina 28804

The involvement of heterotrimeric G proteins in the regulation of adherens junction function is unclear. We identified SNAP as an interactive partner of G₁₂ using yeast two-hybrid screening. glutathione S-transferase pull-down assays showed the selective interaction of SNAP with G₁₂ in COS-7 as well as in human umbilical vein endothelial cells. Using domain swapping experiments, we demonstrated that the N-terminal region of G₁₂ (1–37 amino acids) was necessary and sufficient for its interaction with SNAP. G₁₃ with its N-terminal extension replaced by that of G₁₂ acquired the ability to bind to SNAP, whereas G₁₂ with its N terminus replaced by that of G₁₃ lost this ability. Using four point mutants of SNAP, which alter its ability to bind to the SNARE complex, we determined that the convex rather than the concave surface of SNAP was involved in its interaction with G₁₂. Co-transfection of human umbilical vein endothelial cells with G₁₂ and SNAP stabilized VE-cadherin at the plasma membrane, whereas down-regulation of SNAP with siRNA resulted in the loss of VE-cadherin from the cell surface and, when used in conjunction with G₁₂ overexpression, decreased endothelial barrier function. Our results demonstrate a direct link between the subunit of G₁₂ and SNAP, an essential component of the membrane fusion machinery, and implicate a role for this interaction in regulating the membrane localization of VE-cadherin and endothelial barrier function.

Received for publication, March 15, 2005 , and in revised form, May 24, 2005.

^* This work was supported in part by National Institutes of Health Grants GM56159, GM65160, and HL06078 and by a grant from the American Heart Association (AHA) (to T. V.-Y.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of AHA Predoctoral Fellowship 0310030Z.

^|| To whom correspondence should be addressed: Dept. of Pharmacology (MC 868), University of Illinois, 835 S. Wolcott Ave., Chicago, IL 60612. Tel.: 312-996-9823; Fax: 312-996-1225; E-mail: tvy{at}uic.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. V. Andreeva, M. A. Kutuzov, and T. A. Voyno-Yasenetskaya G{alpha}12 is targeted to the mitochondria and affects mitochondrial morphology and motility FASEB J, August 1, 2008; 22(8): 2821 - 2831. [Abstract] [Full Text] [PDF]

				D. M. Shasby Cell-cell adhesion in lung endothelium Am J Physiol Lung Cell Mol Physiol, March 1, 2007; 292(3): L593 - L607. [Abstract] [Full Text] [PDF]

				A. V. Andreeva, R. Vaiskunaite, M. A. Kutuzov, J. Profirovic, R. A. Skidgel, and T. Voyno-Yasenetskaya Novel Mechanisms of G Protein-Dependent Regulation of Endothelial Nitric-Oxide Synthase Mol. Pharmacol., March 1, 2006; 69(3): 975 - 982. [Abstract] [Full Text] [PDF]

				M. L. Bilodeau and H. E. Hamm Endothelial Nitric-Oxide Synthase Reveals a New Face in G Protein Signaling Mol. Pharmacol., March 1, 2006; 69(3): 677 - 679. [Abstract] [Full Text] [PDF]