p120-Catenin regulates leukocyte transmigration through an effect on VE-cadherin phosphorylation.
Vascular endothelial-cadherin (VE-cad) is localized to adherens junctions at endothelial cell borders and forms a complex with alpha-, beta-, gamma-, and p120-catenins (p120). We previously showed that the VE-cad complex disassociates to form short-lived "gaps" during leukocyte transendothelial migration (TEM); however, whether these gaps are required for leukocyte TEM is not clear. Recently p120 has been shown to control VE-cad surface expression through endocytosis. We hypothesized that p120 regulates VE-cad surface expression, which would in turn have functional consequences for leukocyte transmigration. Here we show that endothelial cells transduced with an adenovirus expressing p120GFP fusion protein significantly increase VE-cad expression. Moreover, endothelial junctions with high p120GFP expression largely prevent VE-cad gap formation and neutrophil leukocyte TEM; if TEM occurs, the length of time required is prolonged. We find no evidence that VE-cad endocytosis plays a role in VE-cad gap formation and instead show that this process is regulated by changes in VE-cad phosphorylation. In fact, a nonphosphorylatable VE-cad mutant prevented TEM. In summary, our studies provide compelling evidence that VE-cad gap formation is required for leukocyte transmigration and identify p120 as a critical intracellular mediator of this process through its regulation of VE-cad expression at junctions.
Alcaide
Pilar
P
Center for Excellence in Vascular Biology, Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.
Newton
Gail
G
Auerbach
Scott
S
Sehrawat
Seema
S
Mayadas
Tanya N
TN
Golan
David E
DE
Yacono
Patrick
P
Vincent
Peter
P
Kowalczyk
Andrew
A
Luscinskas
Francis W
FW
D015703:Antigens, CD / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism*; D051177:Catenins; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D004705:Endocytosis; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004727:Endothelium / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D007962:Leukocytes / Q000166:cytology* / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D000095002:Delta Catenin; D000099152:Cadherin 5
D015703:Antigens, CD; D015820:Cadherins; D051177:Catenins; D015815:Cell Adhesion Molecules; D049452:Green Fluorescent Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D000095002:Delta Catenin; D000099152:Cadherin 5; C000722347:CTNND1 protein, human