GIT1 Mediates Thrombin Signaling in Endothelial Cells
Copyright policy )GIT1 Mediates Thrombin Signaling in Endothelial Cells
Thrombin mediates changes in endothelial barrier function and increases endothelial permeability. A feature of thrombin-enhanced endothelial hyperpermeability is contraction of endothelial cells (ECs), accompanied by formation of focal adhesions (FAs). Recently, a G protein-coupled receptor kinase-interacting protein, GIT1, was shown to regulate FA disassembly. We hypothesized that GIT1 modulates thrombin-induced changes in FAs. In human umbilical vein ECs (HUVECs), thrombin recruited GIT1 to FAs, where GIT1 colocalized with FAK and vinculin. Recruitment of GIT1 to FAs was dependent on activation of the small GTPase RhoA, and Rho kinase, as demonstrated by adenoviral transfection of dominant-negative RhoA and treatment with Y-27632. Thrombin stimulated GIT1 tyrosine phosphorylation with a time course similar to FAK phosphorylation in a Rho kinase- and Src-dependent manner. Depletion of GIT1 with antisense GIT1 oligonucleotides had no effect on basal cell morphology, but increased cell rounding and contraction of HUVECs, increased FA formation, and increased FAK tyrosine phosphorylation in response to thrombin, concomitant with increased endothelial hyperpermeability. These data identify GIT1 as a novel mediator in agonist-dependent signaling in ECs, demonstrate that GIT1 is involved in cell shape changes, and suggest a role for GIT1 as a negative feedback regulator that augments recovery of cell contraction.
- King's College London United Kingdom
- Ludwig Cancer Research United Kingdom
- Delft University of Technology Netherlands
- Kings College London, University of London United Kingdom
- Amsterdam University Medical Centers Netherlands
antisense oligonucleotide, Umbilical Veins, Biomedical Research, cell migration, Pyridines, Cell Cycle Proteins, animal cell, Contractility, Oligodeoxyribonucleotides, Antisense, G protein coupled receptor kinase interacting protein, Transduction, Genetic, Adenovirus, Rho kinase, endothelium cell, enzyme phosphorylation, focal adhesion, RNA, Small Interfering, Enzyme Inhibitors, Phosphorylation, G protein coupled receptor kinase, cellular distribution, Aorta, Cells, Cultured, Feedback, Physiological, depletion, GTPase-Activating Proteins, Focal adhesion kinase, Thrombin, article, guanosine triphosphatase, protein function, Protein-Tyrosine Kinases, thrombin, unclassified drug, rac GTP-Binding Proteins, Protein Transport, priority journal, 4 (1 aminoethyl) n (4 pyridyl)cyclohexanecarboxamide, protein protein interaction, protein transport, RhoA guanine nucleotide binding protein, signal transduction, Signal Transduction, 570, regulatory mechanism, Proto-Oncogene Proteins pp60(c-src), 610, protein localization, Transfection, cell shape, GIT1 protein, Animals, Humans, controlled study, Endothelium, human, Feedback, Biochemical, enzyme substrate, Adaptor Proteins, Signal Transducing, Cell Size, umbilical vein, Focal Adhesions, nonhuman, vinculin, human cell, focal adhesion kinase, Endothelial Cells, protein tyrosine kinase, enzyme activation, negative feedback, Phosphoproteins, Amides, Vinculin, protein phosphorylation, enzyme metabolism, membrane permeability, Focal Adhesion Kinase 1, Focal Adhesion Protein-Tyrosine Kinases, Cattle, genetic transfection, Endothelium, Vascular, rhoA GTP-Binding Protein, Protein Processing, Post-Translational, cell structure
antisense oligonucleotide, Umbilical Veins, Biomedical Research, cell migration, Pyridines, Cell Cycle Proteins, animal cell, Contractility, Oligodeoxyribonucleotides, Antisense, G protein coupled receptor kinase interacting protein, Transduction, Genetic, Adenovirus, Rho kinase, endothelium cell, enzyme phosphorylation, focal adhesion, RNA, Small Interfering, Enzyme Inhibitors, Phosphorylation, G protein coupled receptor kinase, cellular distribution, Aorta, Cells, Cultured, Feedback, Physiological, depletion, GTPase-Activating Proteins, Focal adhesion kinase, Thrombin, article, guanosine triphosphatase, protein function, Protein-Tyrosine Kinases, thrombin, unclassified drug, rac GTP-Binding Proteins, Protein Transport, priority journal, 4 (1 aminoethyl) n (4 pyridyl)cyclohexanecarboxamide, protein protein interaction, protein transport, RhoA guanine nucleotide binding protein, signal transduction, Signal Transduction, 570, regulatory mechanism, Proto-Oncogene Proteins pp60(c-src), 610, protein localization, Transfection, cell shape, GIT1 protein, Animals, Humans, controlled study, Endothelium, human, Feedback, Biochemical, enzyme substrate, Adaptor Proteins, Signal Transducing, Cell Size, umbilical vein, Focal Adhesions, nonhuman, vinculin, human cell, focal adhesion kinase, Endothelial Cells, protein tyrosine kinase, enzyme activation, negative feedback, Phosphoproteins, Amides, Vinculin, protein phosphorylation, enzyme metabolism, membrane permeability, Focal Adhesion Kinase 1, Focal Adhesion Protein-Tyrosine Kinases, Cattle, genetic transfection, Endothelium, Vascular, rhoA GTP-Binding Protein, Protein Processing, Post-Translational, cell structure
citations This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).67 popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.Top 10% influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).Top 10% impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.Top 10%
Citations provided by BIP!Popularity provided by BIP!