TY - JOUR T1 - USP30 sets a trigger threshold for PINK1–PARKIN amplification of mitochondrial ubiquitylation JF - Life Science Alliance JO - Life Sci. Alliance DO - 10.26508/lsa.202000768 VL - 3 IS - 8 SP - e202000768 AU - Emma V Rusilowicz-Jones AU - Jane Jardine AU - Andreas Kallinos AU - Adan Pinto-Fernandez AU - Franziska Guenther AU - Mariacarmela Giurrandino AU - Francesco G Barone AU - Katy McCarron AU - Christopher J Burke AU - Alejandro Murad AU - Aitor Martinez AU - Elena Marcassa AU - Malte Gersch AU - Alexandre J Buckmelter AU - Katherine J Kayser-Bricker AU - Frederic Lamoliatte AU - Akshada Gajbhiye AU - Simon Davis AU - Hannah C Scott AU - Emma Murphy AU - Katherine England AU - Heather Mortiboys AU - David Komander AU - Matthias Trost AU - Benedikt M Kessler AU - Stephanos Ioannidis AU - Michael K Ahlijanian AU - Sylvie Urbé AU - Michael J Clague Y1 - 2020/08/01 UR - https://www.life-science-alliance.org/content/3/8/e202000768.abstract N2 - 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. ER -