Golgi-associated cPLA2alpha regulates endothelial cell-cell junction integrity by controlling the trafficking of transmembrane junction proteins.
In endothelial cells specifically, cPLA2alpha translocates from the cytoplasm to the Golgi complex in response to cell confluence. Considering the link between confluence and cell-cell junction formation, and the emerging role of cPLA2alpha in intracellular trafficking, we tested whether Golgi-associated cPLA2alpha is involved in the trafficking of junction proteins. Here, we show that the redistribution of cPLA2alpha from the cytoplasm to the Golgi correlates with adherens junction maturation and occurs before tight junction formation. Disruption of adherens junctions using a blocking anti-VE-cadherin antibody reverses the association of cPLA2alpha with the Golgi. Silencing of cPLA2alpha and inhibition of cPLA2alpha enzymatic activity using various inhibitors result in the diminished presence of the transmembrane junction proteins VE-cadherin, occludin, and claudin-5 at cell-cell contacts, and in their accumulation at the Golgi. Altogether, our data support the idea that VE-cadherin triggers the relocation of cPLA2alpha to the Golgi and that in turn, Golgi-associated cPLA2alpha regulates the transport of transmembrane junction proteins through or from the Golgi, thereby controlling the integrity of endothelial cell-cell junctions.
Regan-Klapisz
Elsa
E
Cellular Architecture and Dynamics, Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands. E.E.Regan1@uu.nl
Krouwer
Vincent
V
Langelaar-Makkinje
Miriam
M
Nallan
Laxman
L
Gelb
Michael
M
Gerritsen
Hans
H
Verkleij
Arie J
AJ
Post
Jan Andries
JA
D022005:Adherens Junctions / Q000378:metabolism; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D015820:Cadherins / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D062507:Claudin-5; D003593:Cytoplasm / Q000201:enzymology; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006056:Golgi Apparatus / Q000187:drug effects / Q000201:enzymology*; D054513:Group IV Phospholipases A2 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D062793:Occludin; D021381:Protein Transport / Q000187:drug effects; D034622:RNA Interference; D019108:Tight Junctions / Q000378:metabolism*; D000099152:Cadherin 5
D000906:Antibodies; D015703:Antigens, CD; D015820:Cadherins; D062507:Claudin-5; D054513:Group IV Phospholipases A2; D008565:Membrane Proteins; D062793:Occludin; D000099152:Cadherin 5; C107466:CLDN5 protein, human; C568985:OCLN protein, human
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