Global ubiquitylation analysis of mitochondria in primary neurons identifies endogenous Parkin targets following activation of PINK

Output Details

Preprint March 31, 2021

Published November 12, 2021

How activation of PINK1 and Parkin leads to elimination of damaged mitochondria by mitophagy is largely based on cell lines with few studies in neurons. Here, we have undertaken proteomic analysis of mitochondria from mouse neurons to identify ubiquitylated substrates of endogenous Parkin. Comparative analysis with human iNeuron datasets revealed a subset of 49 PINK1 activation–dependent diGLY sites in 22 proteins conserved across mouse and human systems. We use reconstitution assays to demonstrate direct ubiquitylation by Parkin in vitro. We also identified a subset of cytoplasmic proteins recruited to mitochondria that undergo PINK1 and Parkin independent ubiquitylation, indicating the presence of alternate ubiquitin E3 ligase pathways that are activated by mitochondrial depolarization in neurons. Last, we have developed an online resource to search for ubiquitin sites and enzymes in mitochondria of neurons, MitoNUb. These findings will aid future studies to understand Parkin activation in neuronal subtypes. Article published in Science Advances on 12 November 2021. Initial preprint 10.1101/2021.04.01.438131 posted on BioRvix on 01 April 2021.
Identifier (DOI)
10.1126/sciadv.abj0722
Tags
  • Mitochondria
  • Original Research
  • Parkin (PARK2)
  • PINK1
  • Proteomic analysis
  • Ubiquitin

Meet the Authors

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    Odetta Antico

    Key Personnel: Team Alessi

    MRC Protein Phosphorylation and Ubiquitylation Unit

  • Raja Sekhar Nirujogi, PhD

    Key Personnel: Team Alessi

    University of Dundee

  • Miratul Muqit

    Co-PI (Core Leadership): Team Alessi

    University of Dundee

  • J. Wade Harper

    Lead PI (Core Leadership): Team Harper

    Harvard University

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    Alban Ordureau

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    Michael Stevens

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    Francois Singh

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    Marek Gierlinski

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    Erica Barini

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    Mollie L. Rickwood

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    Alan Prescott

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    Rachel Toth

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    Ian G. Ganley