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Site-specific monoubiquitination downregulates Rab5 by disrupting effector binding and guanine nucleotide conversion.

eLife | 2017

Rab GTPases, which are involved in intracellular trafficking pathways, have recently been reported to be ubiquitinated. However, the functions of ubiquitinated Rab proteins remain unexplored. Here we show that Rab5 is monoubiquitinated on K116, K140, and K165. Upon co-transfection with ubiquitin, Rab5 exhibited abnormalities in endosomal localization and EGF-induced EGF receptor degradation. Rab5 K140R and K165R mutants restored these abnormalities, whereas K116R did not. We derived structural models of individual monoubiquitinated Rab5 proteins (mUbRab5s) by solution scattering and observed different conformational flexibilities in a site-specific manner. Structural analysis combined with biochemical data revealed that interactions with downstream effectors were impeded in mUbRab5, whereas GDP release and GTP loading activities were altered in mUbRab5. By contrast, mUbRab5apparently had no effect. We propose a regulatory mechanism of Rab5 where monoubiquitination downregulates effector recruitment and GDP/GTP conversion in a site-specific manner.

Pubmed ID: 28968219 RIS Download

Mesh terms: Cell Line | DNA Mutational Analysis | Down-Regulation | Guanine Nucleotides | Humans | Hydrolysis | Protein Binding | Protein Conformation | Scattering, Small Angle | Ubiquitination | rab5 GTP-Binding Proteins

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