A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1

Ilse Timmerman, Niels Heemskerk, Jeffrey Kroon, Antje Schaefer, Jos van Rijssel, Mark Hoogenboezem, Jakobus van Unen, Joachim Goedhart, Theodorus W J Gadella, Taofei Yin, Yi Wu, Stephan Huveneers, Jaap D van Buul

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    Endothelial cell-cell junctions maintain a restrictive barrier that is tightly regulated to allow dynamic responses to permeability-inducing angiogenic factors, as well as to inflammatory agents and adherent leukocytes. The ability of these stimuli to transiently remodel adherens junctions depends on Rho-GTPase-controlled cytoskeletal rearrangements. How the activity of Rho-GTPases is spatio-temporally controlled at endothelial adherens junctions by guanine-nucleotide exchange factors (GEFs) is incompletely understood. Here, we identify a crucial role for the Rho-GEF Trio in stabilizing junctions based around vascular endothelial (VE)-cadherin (also known as CDH5). Trio interacts with VE-cadherin and locally activates Rac1 at adherens junctions during the formation of nascent contacts, as assessed using a novel FRET-based Rac1 biosensor and biochemical assays. The Rac-GEF domain of Trio is responsible for the remodeling of junctional actin from radial into cortical actin bundles, a crucial step for junction stabilization. This promotes the formation of linear adherens junctions and increases endothelial monolayer resistance. Collectively, our data show the importance of spatio-temporal regulation of the actin cytoskeleton through Trio and Rac1 at VE-cadherin-based cell-cell junctions in the maintenance of the endothelial barrier.

    Original languageEnglish
    Pages (from-to)3041-54
    Number of pages14
    JournalJournal of Cell Science
    Volume128
    Issue number16
    DOIs
    Publication statusPublished - 15 Aug 2015

    Cite this

    Timmerman, I., Heemskerk, N., Kroon, J., Schaefer, A., van Rijssel, J., Hoogenboezem, M., ... van Buul, J. D. (2015). A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1. Journal of Cell Science, 128(16), 3041-54. https://doi.org/10.1242/jcs.168674
    Timmerman, Ilse ; Heemskerk, Niels ; Kroon, Jeffrey ; Schaefer, Antje ; van Rijssel, Jos ; Hoogenboezem, Mark ; van Unen, Jakobus ; Goedhart, Joachim ; Gadella, Theodorus W J ; Yin, Taofei ; Wu, Yi ; Huveneers, Stephan ; van Buul, Jaap D. / A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1. In: Journal of Cell Science. 2015 ; Vol. 128, No. 16. pp. 3041-54.
    @article{9dcc4b31657241aca275eee307cb5822,
    title = "A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1",
    abstract = "Endothelial cell-cell junctions maintain a restrictive barrier that is tightly regulated to allow dynamic responses to permeability-inducing angiogenic factors, as well as to inflammatory agents and adherent leukocytes. The ability of these stimuli to transiently remodel adherens junctions depends on Rho-GTPase-controlled cytoskeletal rearrangements. How the activity of Rho-GTPases is spatio-temporally controlled at endothelial adherens junctions by guanine-nucleotide exchange factors (GEFs) is incompletely understood. Here, we identify a crucial role for the Rho-GEF Trio in stabilizing junctions based around vascular endothelial (VE)-cadherin (also known as CDH5). Trio interacts with VE-cadherin and locally activates Rac1 at adherens junctions during the formation of nascent contacts, as assessed using a novel FRET-based Rac1 biosensor and biochemical assays. The Rac-GEF domain of Trio is responsible for the remodeling of junctional actin from radial into cortical actin bundles, a crucial step for junction stabilization. This promotes the formation of linear adherens junctions and increases endothelial monolayer resistance. Collectively, our data show the importance of spatio-temporal regulation of the actin cytoskeleton through Trio and Rac1 at VE-cadherin-based cell-cell junctions in the maintenance of the endothelial barrier.",
    keywords = "Actin Cytoskeleton/genetics, Antigens, CD/genetics, Cadherins/genetics, Capillary Permeability/genetics, Endothelial Cells/metabolism, Endothelium, Vascular/metabolism, GTP Phosphohydrolases/metabolism, Guanine Nucleotide Exchange Factors/genetics, Human Umbilical Vein Endothelial Cells, Humans, Intercellular Junctions/genetics, Protein-Serine-Threonine Kinases/genetics, Signal Transduction/genetics, rac1 GTP-Binding Protein/genetics",
    author = "Ilse Timmerman and Niels Heemskerk and Jeffrey Kroon and Antje Schaefer and {van Rijssel}, Jos and Mark Hoogenboezem and {van Unen}, Jakobus and Joachim Goedhart and Gadella, {Theodorus W J} and Taofei Yin and Yi Wu and Stephan Huveneers and {van Buul}, {Jaap D}",
    note = "{\circledC} 2015. Published by The Company of Biologists Ltd.",
    year = "2015",
    month = "8",
    day = "15",
    doi = "10.1242/jcs.168674",
    language = "English",
    volume = "128",
    pages = "3041--54",
    journal = "Journal of Cell Science",
    issn = "0021-9533",
    publisher = "Company of Biologists Ltd",
    number = "16",

    }

    Timmerman, I, Heemskerk, N, Kroon, J, Schaefer, A, van Rijssel, J, Hoogenboezem, M, van Unen, J, Goedhart, J, Gadella, TWJ, Yin, T, Wu, Y, Huveneers, S & van Buul, JD 2015, 'A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1' Journal of Cell Science, vol. 128, no. 16, pp. 3041-54. https://doi.org/10.1242/jcs.168674

    A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1. / Timmerman, Ilse; Heemskerk, Niels; Kroon, Jeffrey; Schaefer, Antje; van Rijssel, Jos; Hoogenboezem, Mark; van Unen, Jakobus; Goedhart, Joachim; Gadella, Theodorus W J; Yin, Taofei; Wu, Yi; Huveneers, Stephan; van Buul, Jaap D.

    In: Journal of Cell Science, Vol. 128, No. 16, 15.08.2015, p. 3041-54.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - A local VE-cadherin and Trio-based signaling complex stabilizes endothelial junctions through Rac1

    AU - Timmerman, Ilse

    AU - Heemskerk, Niels

    AU - Kroon, Jeffrey

    AU - Schaefer, Antje

    AU - van Rijssel, Jos

    AU - Hoogenboezem, Mark

    AU - van Unen, Jakobus

    AU - Goedhart, Joachim

    AU - Gadella, Theodorus W J

    AU - Yin, Taofei

    AU - Wu, Yi

    AU - Huveneers, Stephan

    AU - van Buul, Jaap D

    N1 - © 2015. Published by The Company of Biologists Ltd.

    PY - 2015/8/15

    Y1 - 2015/8/15

    N2 - Endothelial cell-cell junctions maintain a restrictive barrier that is tightly regulated to allow dynamic responses to permeability-inducing angiogenic factors, as well as to inflammatory agents and adherent leukocytes. The ability of these stimuli to transiently remodel adherens junctions depends on Rho-GTPase-controlled cytoskeletal rearrangements. How the activity of Rho-GTPases is spatio-temporally controlled at endothelial adherens junctions by guanine-nucleotide exchange factors (GEFs) is incompletely understood. Here, we identify a crucial role for the Rho-GEF Trio in stabilizing junctions based around vascular endothelial (VE)-cadherin (also known as CDH5). Trio interacts with VE-cadherin and locally activates Rac1 at adherens junctions during the formation of nascent contacts, as assessed using a novel FRET-based Rac1 biosensor and biochemical assays. The Rac-GEF domain of Trio is responsible for the remodeling of junctional actin from radial into cortical actin bundles, a crucial step for junction stabilization. This promotes the formation of linear adherens junctions and increases endothelial monolayer resistance. Collectively, our data show the importance of spatio-temporal regulation of the actin cytoskeleton through Trio and Rac1 at VE-cadherin-based cell-cell junctions in the maintenance of the endothelial barrier.

    AB - Endothelial cell-cell junctions maintain a restrictive barrier that is tightly regulated to allow dynamic responses to permeability-inducing angiogenic factors, as well as to inflammatory agents and adherent leukocytes. The ability of these stimuli to transiently remodel adherens junctions depends on Rho-GTPase-controlled cytoskeletal rearrangements. How the activity of Rho-GTPases is spatio-temporally controlled at endothelial adherens junctions by guanine-nucleotide exchange factors (GEFs) is incompletely understood. Here, we identify a crucial role for the Rho-GEF Trio in stabilizing junctions based around vascular endothelial (VE)-cadherin (also known as CDH5). Trio interacts with VE-cadherin and locally activates Rac1 at adherens junctions during the formation of nascent contacts, as assessed using a novel FRET-based Rac1 biosensor and biochemical assays. The Rac-GEF domain of Trio is responsible for the remodeling of junctional actin from radial into cortical actin bundles, a crucial step for junction stabilization. This promotes the formation of linear adherens junctions and increases endothelial monolayer resistance. Collectively, our data show the importance of spatio-temporal regulation of the actin cytoskeleton through Trio and Rac1 at VE-cadherin-based cell-cell junctions in the maintenance of the endothelial barrier.

    KW - Actin Cytoskeleton/genetics

    KW - Antigens, CD/genetics

    KW - Cadherins/genetics

    KW - Capillary Permeability/genetics

    KW - Endothelial Cells/metabolism

    KW - Endothelium, Vascular/metabolism

    KW - GTP Phosphohydrolases/metabolism

    KW - Guanine Nucleotide Exchange Factors/genetics

    KW - Human Umbilical Vein Endothelial Cells

    KW - Humans

    KW - Intercellular Junctions/genetics

    KW - Protein-Serine-Threonine Kinases/genetics

    KW - Signal Transduction/genetics

    KW - rac1 GTP-Binding Protein/genetics

    U2 - 10.1242/jcs.168674

    DO - 10.1242/jcs.168674

    M3 - Article

    VL - 128

    SP - 3041

    EP - 3054

    JO - Journal of Cell Science

    JF - Journal of Cell Science

    SN - 0021-9533

    IS - 16

    ER -