The integral function of the endocytic recycling compartment is regulated by RFFL-mediated ubiquitylation of Rab11 effectors
| Autor: | Sakai, R., Fukuda, R., Unida, S., Aki, M., Ono, Y., Endo, A., Kusumi, S., Koga, D., Fukushima, Toshiaki, Komada, M., Okiyoneda, T. |
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| Rok vydání: | 2018 |
| Předmět: |
Endosome
Endocytic cycle Mutant Vesicular Transport Proteins Endocytic recycling Endosomes Biology Interactome Cell Line 03 medical and health sciences 0302 clinical medicine Ubiquitin Protein Interaction Mapping Humans Rffl 030304 developmental biology Adaptor Proteins Signal Transducing 0303 health sciences Effector Microfilament Proteins Transferrin Ubiquitination Membrane Proteins Biological Transport Cell Biology Endocytosis Cell biology Ubiquitin ligase HEK293 Cells Rab11-FIP rab GTP-Binding Proteins biology.protein Recycling endosome Lysosomes Protein Processing Post-Translational 030217 neurology & neurosurgery HeLa Cells Protein Binding |
| Zdroj: | Journal of cell science. 132(3) |
| ISSN: | 1477-9137 |
| Popis: | Endocytic trafficking is regulated by ubiquitylation (also known as ubiquitination) of cargoes and endocytic machineries. The role of ubiquitylation in lysosomal delivery has been well documented, but its role in the recycling pathway is largely unknown. Here, we report that the ubiquitin (Ub) ligase RFFL regulates ubiquitylation of endocytic recycling regulators. An RFFL dominant-negative (DN) mutant induced clustering of endocytic recycling compartments (ERCs) and delayed endocytic cargo recycling without affecting lysosomal traffic. A BioID RFFL interactome analysis revealed that RFFL interacts with the Rab11 effectors EHD1, MICALL1 and class I Rab11-FIPs. The RFFL DN mutant strongly captured these Rab11 effectors and inhibited their ubiquitylation. The prolonged interaction of RFFL with Rab11 effectors was sufficient to induce the clustered ERC phenotype and to delay cargo recycling. RFFL directly ubiquitylates these Rab11 effectors in vitro, but RFFL knockout (KO) only reduced the ubiquitylation of Rab11-FIP1. RFFL KO had a minimal effect on the ubiquitylation of EHD1, MICALL1, and Rab11-FIP2, and failed to delay transferrin recycling. These results suggest that multiple Ub ligases including RFFL regulate the ubiquitylation of Rab11 effectors, determining the integral function of the ERC. |
| Databáze: | OpenAIRE |
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