Global Landscape and Dynamics of Parkin and USP30-Dependent Ubiquitylomes in iNeurons during Mitophagic Signaling

Ordureau, Alban; Paulo, João A.; Zhang, Jiuchun; An, Heeseon; Swatek, Kirby N.; Cannon, Joe R.; Wan, Qiaoqiao; Komander, David; Harper, J. Wade

doi:10.1016/j.molcel.2019.11.013

Cited by 149 publications

(219 citation statements)

References 80 publications

Supporting

Mentioning

189

Contrasting

Order By: Relevance

“…In our experimental settings, multiple mitochondrial proteins including Miro, mitofusin, milton, and ATP5β are unaltered in dVCP RNAi (Figure 1A and Supplementary Figure S1A), suggesting that mitophagy is intact. Consistent with our results, a very recent paper has shown when VCP is inhibited in iNeurons, the protein abundances of OMM proteins including mitofusin and Miro are not dramatically affected (Ordureau et al, 2020), indicating that VCP does not play a significant role in controlling OMM protein turn over. The different results are likely caused by differences in the experimental systems.…”

Section: Discussionsupporting

confidence: 93%

“…We found no significant changes of multiple mitochondrial markers including fly Miro (DMiro), milton, mitofusin (Marf), and ATP5β in dVCP RNAi mutant flies compared with wild-type controls ( Figures 1A,B and Supplementary Figure S1A). These results are consistent with a recent report showing that in iNeurons with VCP inhibition, there is no drastic change in the abundances of OMM proteins including Miro and mitofusin (Ordureau et al, 2020).…”

Section: Downregulation Of Dvcp Alters Axonal Transport Of Mitochondriasupporting

confidence: 93%

See 1 more Smart Citation

Drosophila VCP/p97 Mediates Dynein-Dependent Retrograde Mitochondrial Motility in Axons

Gonzalez

Wang

2020

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Valosin-containing protein (VCP), also called p97, is an evolutionarily conserved and ubiquitously expressed ATPase with diverse cellular functions. Dominant mutations in VCP are found in a late-onset multisystem degenerative proteinopathy. The neurological manifestations of the disorder include frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). In these patients, long motor neuron axons could be particularly susceptible to defects in axonal transport. However, whether VCP has a physiological function in maintaining axonal transport and whether this role is impaired by disease-causing mutations remains elusive. Here, by employing live-imaging methods in Drosophila larval axons and performing genetic interaction experiments, we discover that VCP regulates the axonal transport of mitochondria. Downregulation of VCP enhances the retrograde transport of mitochondria and reduces the density of mitochondria in larval axons. This unidirectional motility phenotype is rescued by removing one copy of the retrograde motor dynein heavy chain (DHC), or elevating Miro which facilitates anterograde mitochondrial movement by interacting with the anterograde motor kinesin heavy chain (KHC). Importantly, Miro upregulation also significantly improves ATP production of VCP mutant larvae. We investigate human VCP pathogenic mutations in our fly system. We find that expressing these mutations affects mitochondrial transport in the same way as knocking down VCP. Our results reveal a new role of VCP in mediating axonal mitochondrial transport, and provide evidence implicating impaired mitochondrial motility in the pathophysiology of VCP-relevant neurodegenerative diseases.

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Downregulation Of Dvcp Alters Axonal Transport Of Mitochondriasupporting

confidence: 93%

Drosophila VCP/p97 Mediates Dynein-Dependent Retrograde Mitochondrial Motility in Axons

Gonzalez

Wang

2020

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

show abstract

“…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: 65%

“…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%

See 1 more Smart Citation

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

Self Cite

View full text Add to dashboard Cite

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.

show abstract

The proteasome: friend and foe of mitochondrial biogenesis

2020

View full text Add to dashboard Cite

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.

show abstract

Global Landscape and Dynamics of Parkin and USP30-Dependent Ubiquitylomes in iNeurons during Mitophagic Signaling

Cited by 149 publications

References 80 publications

Drosophila VCP/p97 Mediates Dynein-Dependent Retrograde Mitochondrial Motility in Axons

Drosophila VCP/p97 Mediates Dynein-Dependent Retrograde Mitochondrial Motility in Axons

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

The proteasome: friend and foe of mitochondrial biogenesis

Contact Info

Product

Resources

About