Negative Regulation of Autophagy by UBA6-BIRC6–Mediated Ubiquitination of LC3

作者: Rui Jia , Juan S. Bonifacino

DOI: 10.1101/699124

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摘要: Abstract Although the process of autophagy has been extensively studied, mechanisms that regulate it remain insufficiently understood. The ability to manipulate is important not only for addressing fundamental biological questions, but also its possible application treatment various human diseases. To identify novel regulators autophagy, we performed a whole-genome CRISPR/Cas9 knockout screen in H4 neuroblastoma cells gene-edited express endogenous effector LC3B fused tandem GFP and mCherry. Using this methodology, identified ubiquitin-activating (E1) enzyme UBA6 hybrid ubiquitin-conjugating (E2)/ubiquitin-ligase (E3) BIRC6 as regulators. We found these two enzymes cooperate monoubiquitinate on lysine-51, targeting degradation by proteasome. Knockout or increased levels well autophagic flux under conditions nutrient deprivation protein synthesis inhibition. Moreover, depletion KO decreased formation aggresome-like induced structures cells, aggregates an α-synuclein mutant axon rat hippocampal neurons. These findings demonstrate negatively limiting availability LC3B, possibly prevent deleterious effects excessive autophagy. Inhibition BIRC6, other hand, could be used enhance clearance neurodegenerative disorders.

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