Ubiquitylation of lipopolysaccharide by RNF213 during bacterial infection

• Published in: Nature (London) 2021-06, Vol.594 (7861), p.111-116
• Main Authors: Otten, Elsje G., Werner, Emma, Crespillo-Casado, Ana, Boyle, Keith B., Dharamdasani, Vimisha, Pathe, Claudio, Santhanam, Balaji, Randow, Felix
• Format: Article
• Language: English
• Subjects: 13; 14/19; 42/41; 42/89; 631/250/262; 631/326/88; 631/80/39; 631/80/458/582; 82/16; 82/58; 82/83; 96; Adenosine Triphosphatases - metabolism; Animals; Antigens; Antiinfectives and antibacterials; Autophagy; Bacteria; Bacterial diseases; Bacterial infections; Carotid artery; Cell Line; Chromatography; Cytosol; Dynein; Enzymes; Eukaryotes; HeLa Cells; Humanities and Social Sciences; Humans; Infections; Lipid A; Lipids; Lipopolysaccharides; Lipopolysaccharides - immunology; Lipopolysaccharides - metabolism; Membranes; Mice; Molecular weight; multidisciplinary; Phagocytosis; Post-translation; Proteins; RING Finger Domains; Risk analysis; Risk factors; Salmonella; Salmonella Infections - immunology; Salmonella Infections - metabolism; Salmonella Infections - microbiology; Salmonella typhimurium; Science; Science (multidisciplinary); Stenosis; Substrates; Ubiquitin; Ubiquitin - metabolism; Ubiquitin-protein ligase; Ubiquitin-Protein Ligases - metabolism; Ubiquitination
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/s41586-021-03566-4

Ubiquitylation is a widespread post-translational protein modification in eukaryotes and marks bacteria that invade the cytosol as cargo for antibacterial autophagy 1 – 3 . The identity of the ubiquitylated substrate on bacteria is unknown. Here we show that the ubiquitin coat on Salmonella that invade the cytosol is formed through the ubiquitylation of a non-proteinaceous substrate, the lipid A moiety of bacterial lipopolysaccharide (LPS), by the E3 ubiquitin ligase ring finger protein 213 (RNF213). RNF213 is a risk factor for moyamoya disease 4 , 5 , which is a progressive stenosis of the supraclinoid internal carotid artery that causes stroke (especially in children) 6 , 7 . RNF213 restricts the proliferation of cytosolic Salmonella and is essential for the generation of the bacterial ubiquitin coat, both directly (through the ubiquitylation of LPS) and indirectly (through the recruitment of LUBAC, which is a downstream E3 ligase that adds M1-linked ubiquitin chains onto pre-existing ubiquitin coats 8 ). In cells that lack RNF213, bacteria do not attract ubiquitin-dependent autophagy receptors or induce antibacterial autophagy. The ubiquitylation of LPS on Salmonella that invade the cytosol requires the dynein-like core of RNF213, but not its RING domain. Instead, ubiquitylation of LPS relies on an RZ finger in the E3 shell. We conclude that ubiquitylation extends beyond protein substrates and that ubiquitylation of LPS triggers cell-autonomous immunity, and we postulate that non-proteinaceous substances other than LPS may also become ubiquitylated. Upon Salmonella invasion of the mammalian cytosol, ubiquitylation of a non-proteinaceous substrate—the lipid A moiety of bacterial lipopolysaccharide—by the E3 ubiquitin ligase RNF213 marks the bacteria as cargo for antibacterial autophagy.