Desmoplakin expression and organization at human umbilical vein endothelial cell-to-cell junctions.
Desmoplakin is an intracellular component of desmosomes which plays a role in the anchorage of intermediate filaments to these structures. We report here that, despite the absence of desmosomes, cultured endothelial cells from human umbilical vein express desmoplakin I and II both at mRNA and protein level. Desmoplakin I/II are found only in the detergent insoluble fraction suggesting that most of the protein is linked to the cytoskeleton. Desmoplakin I/II could be detected by western blot only in long confluent cells even if desmoplakin mRNA levels are unchanged by cell confluency. This suggests that desmoplakin might be stabilized at protein level by its association with junctional components. Immunofluorescence confocal microscopy showed that desmoplakin codistributes with VE-cadherin and plakoglobin along the lateral cell membrane. In contrast, desmoplakin localization was distinct from that of PECAM, an endothelial specific junctional protein localized outside adherence junctions. Endothelial cells do not have keratins but they express vimentin. In confluent cells vimentin forms peripheral filaments which attach to the cell membrane in areas at desmoplakin localization. These data suggest that desmoplakin may participate in the molecular organization of interendothelial junctions by interacting with VE-cadherin and promoting vimentin anchorage. This new type of intercellular junction seems to correspond to the "complexus adhaerentes' described in vivo in lymphatic endothelium.
Valiron
O
O
CEA, Laboratoire d'Hematologie, INSERM U217, Grenoble, France.
Chevrier
V
V
Usson
Y
Y
Breviario
F
F
Job
D
D
Dejana
E
E
D015152:Blotting, Northern; D015815:Cell Adhesion Molecules / Q000096:biosynthesis*; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000096:biosynthesis* / Q000235:genetics; D051180:Desmoplakins; D003896:Desmosomes / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007365:Intercellular Junctions / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology* / Q000378:metabolism; D051185:gamma Catenin
D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; C495294:DSP protein, human; D051180:Desmoplakins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger; D051185:gamma Catenin