Ubiquitination and receptor-mediated mitophagy converge to eliminate oxidation-damaged mitochondria during hypoxia

The contribution of the Ubiquitin-Proteasome System (UPS) to mitophagy has been largely attributed to the E3 ubiquitin ligase Parkin. Here we show that in response to the oxidative stress associated with hypoxia or the hypoxia mimic CoCl2, the damaged and fragmented mitochondria are removed by Parkin-independent mitophagy. Mitochondria isolated from hypoxia or CoCl2-treated cells exhibited extensive ubiquitination, predominantly Lysine 48-linked and involves the degradation of key mitochondrial proteins such as the mitofusins MFN1/2, or the import channel component TOM20. Reflecting the critical role of mitochondrial protein degradation, pmteasome inhibition blocked CoCl2-induced mitophagy. The five conserved ubiquitin-binding autophagy receptors (p62, NDP52, Optineurin, NBR1, TAX1BP1) were dispensable for the ensuing mitophagy, suggesting that the mitophagy step itself was independent of ubiquitination. Instead, the expression of two ubiquitin-independent mitophagy receptor proteins BNIP3 and NIX was induced by hypoxia or CoCl2-treatment followed by their recruitment to the oxidation-damaged mitochondria. By employing BNIP3/NIX double knockout and DRP1-null cell lines, we confirmed that mitochondrial clearance relies on DRP1-dependent mitochondrial fragmentation and BNIP3/NIX-mediated mitophagy. General antioxidants such as N-Acetyl Cysteine (NAC) or the mitochondria-specific Mitoquinone prevented HIF-1 alpha stabilization, ameliorated hypoxiarelated mitochondrial oxidative stress, and suppressed mitophagy. We conclude that the UPS and receptormediated autophagy converge to eliminate oxidation-damaged mitochondria.

Automated summary

The study reveals that damaged mitochondria can be cleared via Parkin-independent mitophagy under oxidative stress conditions induced by hypoxia or CoCl2. The process involves extensive ubiquitination and degradation of key mitochondrial proteins, independent of the five ubiquitin-binding autophagy receptors. Additionally, the clearance of oxidation-damaged mitochondria relies on DRP1-mediated mitochondrial fragmentation and BNIP3/NIX-mediated mitophagy.