Dynamic Regulation of Mitochondrial Import by the Ubiquitin System

Phu, Lilian; Rose, Christopher M.; Tea, Joy S.; Wall, Christopher E.; Verschueren, Erik; Cheung, Tommy K.; Kirkpatrick, Donald S.; Bingol, Baris

doi:10.1016/j.molcel.2020.02.012

Cited by 115 publications

(144 citation statements)

References 90 publications

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“…In fact, in a recently published whole genome screen for mitophagy regulators in Parkin overexpressing C2C12 myoblasts, USP30 is the most prominent mitochondrial annotated negative regulator (25). Our study contributes to a body of evidence that translates these finding to systems with endogenous Parkin expression levels (20,56,57). The physiological defects associated with PINK1/Parkin loss of function in Parkinson's Disease are likely to accumulate slowly.…”

Section: Discussionmentioning

confidence: 66%

“…Ordureau et al generate iNeurons and examine the impact of USP30 genetic loss (56). Phu et al use HEK293 cells to compare genetic loss with a USP30 inhibitor that is related to the one we describe here (57). Note that, in that latter study a much higher concentration of inhibitor has been used (5µM vs 200nM).…”

Section: Discussionmentioning

confidence: 98%

“…Whilst our manuscript was in preparation, two complementary studies have been published that also highlight the centrality of the TOM complex to USP30 function (56,57). All three studies use global proteome and ubiquitylome profiling.…”

Section: Discussionmentioning

confidence: 99%

“…Although ubiquitylation can occur within mitochondria (58), USP30 is an outer mitochondrial membrane protein whose catalytic activity is facing towards the cytosol (18,20). Hence, it has been suggested that this reflects ubiquitylation of newly synthesised proteins engaging with the TOM complex (56,57). Thus USP30 might sit at the gate of the import complex pore and strip off ubiquitin as a prerequisite for entry.…”

Section: Discussionmentioning

confidence: 99%

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A novel USP30 inhibitor recapitulates genetic loss of USP30 and sets the trigger for PINK1-PARKIN amplification of mitochondrial ubiquitylation

Rusilowicz‐Jones

Jardine

Pinto-Fernández

et al. 2020

Preprint

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The mitochondrial deubiquitylase USP30 negatively regulates the selective autophagy of damaged mitochondria. It has been proposed as an actionable target to alleviate the loss of function of the mitophagy pathway governed by the Parkinson's Disease associated genes PINK1 and PRKN. We present the characterisation of a N-cyano pyrrolidine derived compound, FT3967385, with high selectivity for USP30. The compound is well tolerated with no loss of total mitochondrial mass. We demonstrate that ubiquitylation of TOM20, a component of the outer mitochondrial membrane import machinery that directly interacts with USP30, represents a robust biomarker for both USP30 loss and inhibition. We have conducted proteomics analyses on a SHSY5Y neuroblastoma cell line model to directly compare the effects of genetic loss of USP30 with selective inhibition in an unbiased fashion. 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 most 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. In this model, 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 and to a greater extent in cells either lacking USP30 or subject to USP30 inhibition.

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Section: Discussionmentioning

confidence: 66%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

A novel USP30 inhibitor recapitulates genetic loss of USP30 and sets the trigger for PINK1-PARKIN amplification of mitochondrial ubiquitylation

Rusilowicz‐Jones

Jardine

Pinto-Fernández

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…While our manuscript was in preparation, two complementary studies have been published that also highlight the centrality of the TOM complex to USP30 function (Ordureau et al, 2020;Phu et al, 2020). All three studies use global proteome and ubiquitylome profiling.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2020

View full text Add to dashboard Cite

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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The proteasome: friend and foe of mitochondrial biogenesis

2020

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Most mitochondrial proteins are synthesized in the cytosol and subsequently translocated as unfolded polypeptides into mitochondria. Cytosolic chaperones maintain precursor proteins in an import‐competent state. This post‐translational import reaction is under surveillance of the cytosolic ubiquitin‐proteasome system, which carries out several distinguishable activities. On the one hand, the proteasome degrades nonproductive protein precursors from the cytosol and nucleus, import intermediates that are stuck in mitochondrial translocases, and misfolded or damaged proteins from the outer membrane and the intermembrane space. These surveillance activities of the proteasome are essential for mitochondrial functionality, as well as cellular fitness and survival. On the other hand, the proteasome competes with mitochondria for nonimported cytosolic precursor proteins, which can compromise mitochondrial biogenesis. In order to balance the positive and negative effects of the cytosolic protein quality control system on mitochondria, mitochondrial import efficiency directly regulates the capacity of the proteasome via transcription factor Rpn4 in yeast and nuclear respiratory factor (Nrf) 1 and 2 in animal cells. In this review, we provide a thorough overview of how the proteasome regulates mitochondrial biogenesis.

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Dynamic Regulation of Mitochondrial Import by the Ubiquitin System

Abstract: Highlights d USP30 and March5 reciprocally modify import substrates at the TOM complex d USP30 promotes mitochondrial import of ubiquitinated substrates d March5 directs degradation of import intermediates by proteasomes d USP30 KO mice have reduced abundance of the TOM complex

Cited by 115 publications

References 90 publications

A novel USP30 inhibitor recapitulates genetic loss of USP30 and sets the trigger for PINK1-PARKIN amplification of mitochondrial ubiquitylation

A novel USP30 inhibitor recapitulates genetic loss of USP30 and sets the trigger for PINK1-PARKIN amplification of mitochondrial ubiquitylation

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

The proteasome: friend and foe of mitochondrial biogenesis

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