ZO-1 controls endothelial adherens junctions, cell-cell tension, angiogenesis, and barrier formation.
Intercellular junctions are crucial for mechanotransduction, but whether tight junctions contribute to the regulation of cell-cell tension and adherens junctions is unknown. Here, we demonstrate that the tight junction protein ZO-1 regulates tension acting on VE-cadherin-based adherens junctions, cell migration, and barrier formation of primary endothelial cells, as well as angiogenesis in vitro and in vivo. ZO-1 depletion led to tight junction disruption, redistribution of active myosin II from junctions to stress fibers, reduced tension on VE-cadherin and loss of junctional mechanotransducers such as vinculin and PAK2, and induced vinculin dissociation from the α-catenin-VE-cadherin complex. Claudin-5 depletion only mimicked ZO-1 effects on barrier formation, whereas the effects on mechanotransducers were rescued by inhibition of ROCK and phenocopied by JAM-A, JACOP, or p114RhoGEF down-regulation. ZO-1 was required for junctional recruitment of JACOP, which, in turn, recruited p114RhoGEF. ZO-1 is thus a central regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis, and barrier formation.
Department of Cell Biology, UCL Institute of Ophthalmology, University College London, London EC1V 9EL, England, UK.
Olga
O
Tornavaca
Department of Cell Biology, UCL Institute of Ophthalmology, University College London, London EC1V 9EL, England, UK.
Minghao
M
Chia
National Heart and Lung Institute (NHLI) Vascular Sciences Unit, Imperial Centre for Translational and Experimental Medicine (ICTEM), Hammersmith Hospital, Imperial College London, London W12 0NN, England, UK.
Neil
N
Dufton
National Heart and Lung Institute (NHLI) Vascular Sciences Unit, Imperial Centre for Translational and Experimental Medicine (ICTEM), Hammersmith Hospital, Imperial College London, London W12 0NN, England, UK.
Lourdes Osuna
LO
Almagro
Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA 23284.
Daniel E
DE
Conway
National Heart and Lung Institute (NHLI) Vascular Sciences Unit, Imperial Centre for Translational and Experimental Medicine (ICTEM), Hammersmith Hospital, Imperial College London, London W12 0NN, England, UK.
Anna M
AM
Randi
Department of Medicine and Department of Cell Biology, Yale University, New Haven, CT 06520 Department of Medicine and Department of Cell Biology, Yale University, New Haven, CT 06520.
Martin A
MA
Schwartz
Department of Cell Biology, UCL Institute of Ophthalmology, University College London, London EC1V 9EL, England, UK.
Karl
K
Matter
Department of Cell Biology, UCL Institute of Ophthalmology, University College London, London EC1V 9EL, England, UK m.balda@ucl.ac.uk.
Maria S
MS
Balda
D000205:Actomyosin / Q000378:metabolism; D022005:Adherens Junctions / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002199:Capillary Permeability; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002465:Cell Movement; D002478:Cells, Cultured; D062507:Claudin-5 / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D042783:Endothelial Cells / Q000502:physiology*; D006801:Humans; D040542:Mechanotransduction, Cellular; D008810:Mice, Inbred C57BL; D009218:Myosins / Q000378:metabolism; D018919:Neovascularization, Physiologic; D021381:Protein Transport; D011956:Receptors, Cell Surface / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000502:physiology*; D000099152:Cadherin 5
D000205:Actomyosin; D015703:Antigens, CD; D015820:Cadherins; D015815:Cell Adhesion Molecules; D062507:Claudin-5; D003598:Cytoskeletal Proteins; D009218:Myosins; D011956:Receptors, Cell Surface; D062826:Zonula Occludens-1 Protein; D000099152:Cadherin 5; C533277:CGNL1 protein, human; C107466:CLDN5 protein, human; C407243:F11R protein, human; C569042:TJP1 protein, human