JRAB/MICAL-L2 Is a Junctional Rab13-binding Protein Mediating the Endocytic Recycling of Occludin
Open Access
- 1 May 2006
- journal article
- Published by American Society for Cell Biology (ASCB) in Molecular Biology of the Cell
- Vol. 17 (5), 2465-2475
- https://doi.org/10.1091/mbc.e05-09-0826
Abstract
The dynamic turnover of tight junctions (TJs) is essential for epithelial-mesenchymal transitions and/or mesenchymal-epithelial transitions during epithelial morphogenesis. We previously demonstrated that Rab13 specifically mediates the endocytic recycling of occludin. Here, we identified MICAL-L2 (molecule interacting with CasL-like 2) as a novel Rab13-binding protein. Immunoprecipitation and immunofluorescence microscopy showed that MICAL-L2 specifically bound to the GTP-bound form of Rab13 via its C terminus, which contained a coiled-coil domain, and localized at TJs in epithelial MTD-1A cells. Recycling assay demonstrated that a MICAL-L2 mutant lacking the Rab13-binding domain (MICAL-L2-N) specifically inhibited the endocytic recycling of occludin but not transferrin receptor. Ca2+ switch assay further revealed that MICAL-L2-N as well as Rab13 Q67L inhibited the recruitment of occludin to the plasma membrane, the development of transepithelial electrical resistance, and the formation of a paracellular diffusion barrier. MICAL-L2 was displaced from TJs upon actin depolymerization and was distributed along radiating actin cables and stress fibers in Ca2+-depleted MTD-1A and fibroblastic NIH3T3 cells, respectively. These results suggest that MICAL-L2 mediates the endocytic recycling of occludin and the formation of functional TJs by linking Rab13 to actin cytoskeleton. We rename MICAL-L2 as JRAB (junctional Rab13-binding protein).Keywords
This publication has 57 references indexed in Scilit:
- Mechanism of IFN-γ-induced Endocytosis of Tight Junction Proteins: Myosin II-dependent Vacuolarization of the Apical Plasma MembraneMolecular Biology of the Cell, 2005
- Knockdown of occludin expression leads to diverse phenotypic alterations in epithelial cellsAmerican Journal of Physiology-Cell Physiology, 2005
- The MICAL proteins and rab1: a possible link to the cytoskeleton?Biochemical and Biophysical Research Communications, 2005
- JACOP, a Novel Plaque Protein Localizing at the Apical Junctional Complex with Sequence Similarity to CingulinPublished by Elsevier BV ,2004
- Role for Actin Filament Turnover and a Myosin II Motor in Cytoskeleton-driven Disassembly of the Epithelial Apical Junctional ComplexMolecular Biology of the Cell, 2004
- The tight junction: a multifunctional complexAmerican Journal of Physiology-Cell Physiology, 2004
- Platelet‐derived growth factor mediates tight junction redistribution and increases permeability in MDCK cellsJournal of Cellular Physiology, 2002
- A Family of Rab27-binding ProteinsJournal of Biological Chemistry, 2002
- Physical and Functional Interaction of Rabphilin-3A with α-ActininPublished by Elsevier BV ,1996
- Occludin: a novel integral membrane protein localizing at tight junctions.The Journal of cell biology, 1993