2. Academic Validation
  3. USP30 sets a trigger threshold for PINK1-PARKIN amplification of mitochondrial ubiquitylation

USP30 sets a trigger threshold for PINK1-PARKIN amplification of mitochondrial ubiquitylation

  • Life Sci Alliance. 2020 Jul 7;3(8):e202000768. doi: 10.26508/lsa.202000768.
Emma V Rusilowicz-Jones 1,  Jane Jardine 1,  Andreas Kallinos 1,  Adan Pinto-Fernandez 2,  Franziska Guenther 3,  Mariacarmela Giurrandino 3,  Francesco G Barone 1,  Katy McCarron 1,  Christopher J Burke 4,  Alejandro Murad 4,  Aitor Martinez 1,  Elena Marcassa 1,  Malte Gersch 5 6,  Alexandre J Buckmelter 4,  Katherine J Kayser-Bricker 4,  Frederic Lamoliatte 7,  Akshada Gajbhiye 7,  Simon Davis 2,  Hannah C Scott 2,  Emma Murphy 3,  Katherine England 3,  Heather Mortiboys 8,  David Komander 9 10,  Matthias Trost 7,  Benedikt M Kessler 2,  Stephanos Ioannidis 4,  Michael K Ahlijanian 4,  Sylvie Urbé 11,  Michael J Clague 12
Affiliations

Affiliations

  • 1 Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, UK.
  • 2 Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
  • 3 Alzheimer's Research UK, Oxford Drug Discovery Institute, Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
  • 4 FORMA Therapeutics, Watertown, MA, USA.
  • 5 Chemical Genomics Centre, Max-Planck-Institute of Molecular Physiology, Dortmund, Germany.
  • 6 Department of Chemistry and Chemical Biology, Technische Universität Dortmund, Dortmund, Germany.
  • 7 Laboratory for Biological Mass Spectrometry, Newcastle University Biosciences Institute, Faculty of Medical Sciences, University of Newcastle, Newcastle, UK.
  • 8 Sheffield Institute for Translational Neuroscience (SITraN), University of Sheffield, Sheffield, UK.
  • 9 Ubiquitin Signalling Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.
  • 10 Department of Medical Biology, University of Melbourne, Melbourne, Australia.
  • 11 Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, UK [email protected].
  • 12 Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, UK [email protected].
PMID: 32636217 DOI: 10.26508/lsa.202000768
Abstract

The mitochondrial deubiquitylase USP30 negatively regulates the selective Autophagy of damaged mitochondria. We present the characterisation of an N-cyano pyrrolidine compound, FT3967385, with high selectivity for USP30. We demonstrate that ubiquitylation of TOM20, a component of the outer mitochondrial membrane import machinery, represents a robust biomarker for both USP30 loss and inhibition. A proteomics analysis, on a SHSY5Y neuroblastoma cell line model, directly compares the effects of genetic loss of USP30 with chemical inhibition. We have thereby identified a subset of ubiquitylation events consequent to mitochondrial depolarisation that are USP30 sensitive. Within responsive elements of the ubiquitylome, several components of the outer mitochondrial membrane transport (TOM) complex are prominent. Thus, our data support a model whereby USP30 can regulate the availability of ubiquitin at the specific site of mitochondrial PINK1 accumulation following membrane depolarisation. USP30 deubiquitylation of TOM complex components dampens the trigger for the Parkin-dependent amplification of mitochondrial ubiquitylation leading to Mitophagy. Accordingly, PINK1 generation of phospho-Ser65 ubiquitin proceeds more rapidly in cells either lacking USP30 or subject to USP30 inhibition.

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