title: USP30 and parkin homeostatically regulate atypical ubiquitin chains on mitochondria.
abstract: Multiple lines of evidence indicate that mitochondrial dysfunction is central to Parkinson's disease. Here we investigate the mechanism by which parkin, an E3 ubiquitin ligase, and USP30, a mitochondrion-localized deubiquitylase, regulate mitophagy. We find that mitochondrial damage stimulates parkin to assemble Lys 6, Lys 11 and Lys 63 chains on mitochondria, and that USP30 is a ubiquitin-specific deubiquitylase with a strong preference for cleaving Lys 6- and Lys 11-linked multimers. Using mass spectrometry, we show that recombinant USP30 preferentially removes these linkage types from intact ubiquitylated mitochondria and counteracts parkin-mediated ubiquitin chain formation in cells. These results, combined with a series of chimaera and localization studies, afford insights into the mechanism by which a balance of ubiquitylation and deubiquitylation regulates mitochondrial homeostasis, and suggest a general mechanism for organelle autophagy.
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Christian N
authors.initials: CN
authors.lastname: Cunningham
authors.affiliation: Department of Protein Chemistry, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Joshua M
authors.initials: JM
authors.lastname: Baughman
authors.affiliation: Department of Protein Chemistry, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Lilian
authors.initials: L
authors.lastname: Phu
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Joy S
authors.initials: JS
authors.lastname: Tea
authors.affiliation: Department of Structural Biology, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Yu
authors.affiliation: Department of Structural Biology, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Mary
authors.initials: M
authors.lastname: Coons
authors.affiliation: Department of Protein Chemistry, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Donald S
authors.initials: DS
authors.lastname: Kirkpatrick
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Baris
authors.initials: B
authors.lastname: Bingol
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., 1 DNA Way South San Francisco, California 94080, USA.
authors.forename: Jacob E
authors.initials: JE
authors.lastname: Corn
mesh_terms: D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D020134:Catalytic Domain; D002457:Cell Extracts; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006706:Homeostasis / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D008239:Lysine / Q000378:metabolism; D013058:Mass Spectrometry; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000378:metabolism*; D063306:Mitophagy / Q000187:drug effects; D008954:Models, Biological; D020675:Peroxisomes / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity / Q000187:drug effects; D013869:Thiolester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D064570:Ubiquitin-Specific Proteases / Q000378:metabolism; D054875:Ubiquitination / Q000187:drug effects
chemical_list: D002457:Cell Extracts; D024101:Mitochondrial Proteins; D025801:Ubiquitin; C532819:Usp30 protein, human; D049452:Green Fluorescent Proteins; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D013869:Thiolester Hydrolases; D064570:Ubiquitin-Specific Proteases; D008239:Lysine