Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells
The mechanisms by which the microtubule cytoskeleton regulates the permeability of endothelial barrier are not well understood. Here, we demonstrate that microtubule-associated end-binding protein 3 (EB3), a core component of the microtubule plus-end protein complex, binds to inositol 1,4,5-trisphos...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2015-07-01
|
| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124715005847 |
| _version_ | 1818534615474765824 |
|---|---|
| author | Melissa Geyer Fei Huang Ying Sun Stephen M. Vogel Asrar B. Malik Colin W. Taylor Yulia A. Komarova |
| author_facet | Melissa Geyer Fei Huang Ying Sun Stephen M. Vogel Asrar B. Malik Colin W. Taylor Yulia A. Komarova |
| author_sort | Melissa Geyer |
| collection | DOAJ |
| description | The mechanisms by which the microtubule cytoskeleton regulates the permeability of endothelial barrier are not well understood. Here, we demonstrate that microtubule-associated end-binding protein 3 (EB3), a core component of the microtubule plus-end protein complex, binds to inositol 1,4,5-trisphosphate receptors (IP3Rs) through an S/TxIP EB-binding motif. In endothelial cells, α-thrombin, a pro-inflammatory mediator that stimulates phospholipase Cβ, increases the cytosolic Ca2+ concentration and elicits clustering of IP3R3s. These responses, and the resulting Ca2+-dependent phosphorylation of myosin light chain, are prevented by depletion of either EB3 or mutation of the TxIP motif of IP3R3 responsible for mediating its binding to EB3. We also show that selective EB3 gene deletion in endothelial cells of mice abrogates α-thrombin-induced increase in endothelial permeability. We conclude that the EB3-mediated interaction of IP3Rs with microtubules controls the assembly of IP3Rs into effective Ca2+ signaling clusters, which thereby regulate microtubule-dependent endothelial permeability. |
| first_indexed | 2024-12-11T18:13:52Z |
| format | Article |
| id | doaj.art-d6964fc92987402598060f2ce2003146 |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-12-11T18:13:52Z |
| publishDate | 2015-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-d6964fc92987402598060f2ce20031462022-12-22T00:55:29ZengElsevierCell Reports2211-12472015-07-01121798910.1016/j.celrep.2015.06.001Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial CellsMelissa Geyer0Fei Huang1Ying Sun2Stephen M. Vogel3Asrar B. Malik4Colin W. Taylor5Yulia A. Komarova6Department of Pharmacology and The Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL 60612, USADepartment of Pharmacology and The Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL 60612, USADepartment of Pharmacology and The Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL 60612, USADepartment of Pharmacology and The Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL 60612, USADepartment of Pharmacology and The Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL 60612, USADepartment of Pharmacology, University of Cambridge, Cambridge CB2 1PD, UKDepartment of Pharmacology and The Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL 60612, USAThe mechanisms by which the microtubule cytoskeleton regulates the permeability of endothelial barrier are not well understood. Here, we demonstrate that microtubule-associated end-binding protein 3 (EB3), a core component of the microtubule plus-end protein complex, binds to inositol 1,4,5-trisphosphate receptors (IP3Rs) through an S/TxIP EB-binding motif. In endothelial cells, α-thrombin, a pro-inflammatory mediator that stimulates phospholipase Cβ, increases the cytosolic Ca2+ concentration and elicits clustering of IP3R3s. These responses, and the resulting Ca2+-dependent phosphorylation of myosin light chain, are prevented by depletion of either EB3 or mutation of the TxIP motif of IP3R3 responsible for mediating its binding to EB3. We also show that selective EB3 gene deletion in endothelial cells of mice abrogates α-thrombin-induced increase in endothelial permeability. We conclude that the EB3-mediated interaction of IP3Rs with microtubules controls the assembly of IP3Rs into effective Ca2+ signaling clusters, which thereby regulate microtubule-dependent endothelial permeability.http://www.sciencedirect.com/science/article/pii/S2211124715005847 |
| spellingShingle | Melissa Geyer Fei Huang Ying Sun Stephen M. Vogel Asrar B. Malik Colin W. Taylor Yulia A. Komarova Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells Cell Reports |
| title | Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells |
| title_full | Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells |
| title_fullStr | Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells |
| title_full_unstemmed | Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells |
| title_short | Microtubule-Associated Protein EB3 Regulates IP3 Receptor Clustering and Ca2+ Signaling in Endothelial Cells |
| title_sort | microtubule associated protein eb3 regulates ip3 receptor clustering and ca2 signaling in endothelial cells |
| url | http://www.sciencedirect.com/science/article/pii/S2211124715005847 |
| work_keys_str_mv | AT melissageyer microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells AT feihuang microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells AT yingsun microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells AT stephenmvogel microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells AT asrarbmalik microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells AT colinwtaylor microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells AT yuliaakomarova microtubuleassociatedproteineb3regulatesip3receptorclusteringandca2signalinginendothelialcells |