The ubiquitin isopeptidase USP10 deubiquitinates LC3B to increase LC3B levels and autophagic activity

• Published in: The Journal of biological chemistry Vol. 296; p. 100405
• Main Authors: Jia, Rui, Bonifacino, Juan S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2021; American Society for Biochemistry and Molecular Biology
• Subjects: autophagy; Autophagy - physiology; Cell Line; Cell Line, Tumor; CRISPR/cas; deubiquitination; Endopeptidases - metabolism; Humans; Inhibitor of Apoptosis Proteins; Intracellular Signaling Peptides and Proteins; LC3; Microtubule-Associated Proteins - metabolism; Microtubule-Associated Proteins - physiology; protein aggregation; Protein Processing, Post-Translational; Sequestosome-1 Protein; ubiquitin; Ubiquitin Thiolesterase - genetics; Ubiquitin Thiolesterase - metabolism; Ubiquitin Thiolesterase - physiology; Ubiquitin-Activating Enzymes - metabolism; Ubiquitination; USP10; MAGeCK; RRA; autophagy; KO; USP10; LC3B; ALIS; tfLC3B; ubiquitin; DUB; CHX; ATG; protein aggregation; KD; deubiquitination; NGS; CRISPR/cas; LC3
• DOI: 10.1016/j.jbc.2021.100405
• ISSN: 0021-9258; 1083-351X

Components of the autophagy machinery are subject to regulation by various posttranslational modifications. Previous studies showed that monoubiquitination of LC3B catalyzed by the ubiquitin-activating enzyme UBA6 and ubiquitin-conjugating enzyme/ubiquitin ligase BIRC6 targets LC3B for proteasomal degradation, thus reducing LC3B levels and autophagic activity under conditions of stress. However, mechanisms capable of counteracting this process are not known. Herein, we report that LC3B ubiquitination is reversed by the action of the deubiquitinating enzyme USP10. We identified USP10 in a CRISPR-Cas9 knockout screen for ubiquitination-related genes that regulate LC3B levels. Biochemical analyses showed that silencing of USP10 reduces the levels of both the LC3B-I and LC3B-II forms of LC3B through increased ubiquitination and proteasomal degradation. In turn, the reduced LC3B levels result in slower degradation of the autophagy receptors SQSTM1 and NBR1 and an increased accumulation of puromycin-induced aggresome-like structures. Taken together, these findings indicate that the levels of LC3B and autophagic activity are controlled through cycles of LC3B ubiquitination and deubiquitination.