Mitochondrial Bol1 and Bol3 function as assembly factors for specific iron-sulfur proteins

  1. Marta A Uzarska
  2. Veronica Nasta
  3. Benjamin D Weiler
  4. Farah Spantgar
  5. Simone Ciofi-Baffoni
  6. Maria Rosaria Saviello
  7. Leonardo Gonnelli
  8. Ulrich Mühlenhoff
  9. Lucia Banci  Is a corresponding author
  10. Roland Lill  Is a corresponding author
  1. Philipps-Universität, Germany
  2. University of Florence, Italy
  3. Philipps-Universität Marburg, Germany

Abstract

Assembly of mitochondrial iron-sulfur (Fe/S) proteins is a key process of cells, and defects cause many rare diseases. In the first phase of this pathway, ten Fe/S cluster (ISC) assembly components synthesize and insert [2Fe-2S] clusters. The second phase is dedicated to the assembly of [4Fe-4S] proteins, yet this part is poorly understood. Here, we characterize the BOLA family proteins Bol1 and Bol3 as specific mitochondrial ISC assembly factors that facilitate [4Fe-4S] cluster insertion into a subset of mitochondrial proteins such as lipoate synthase and succinate dehydrogenase. Bol1-Bol3 perform largely overlapping functions, yet cannot replace the ISC protein Nfu1 that also participates in this phase of Fe/S protein biogenesis. Bol1 and Bol3 form dimeric complexes with both monothiol glutaredoxin Grx5 and Nfu1. Complex formation differentially influences the stability of the Grx5-Bol-shared Fe/S clusters. Our findings provide the biochemical basis for explaining the pathological phenotypes of patients with mutations in BOLA3.

Article and author information

Author details

  1. Marta A Uzarska

    Institut für Zytobiologie und Zytopathologie, Philipps-Universität, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Veronica Nasta

    Magnetic Resonance Center CERM, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  3. Benjamin D Weiler

    Institut für Zytobiologie, Philipps-Universität Marburg, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Farah Spantgar

    Institut für Zytobiologie und Zytopathologie, Philipps-Universität, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Simone Ciofi-Baffoni

    Magnetic Resonance Center CERM, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  6. Maria Rosaria Saviello

    Magnetic Resonance Center CERM, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  7. Leonardo Gonnelli

    Magnetic Resonance Center CERM, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  8. Ulrich Mühlenhoff

    Institut für Zytobiologie und Zytopathologie, Philipps-Universität, Marburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Lucia Banci

    Magnetic Resonance Center CERM, University of Florence, Florence, Italy
    For correspondence
    banci@cerm.unifi.it
    Competing interests
    The authors declare that no competing interests exist.
  10. Roland Lill

    Institut für Zytobiologie, Philipps-Universität Marburg, Marburg, Germany
    For correspondence
    lill@staff.uni-marburg.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8345-6518

Funding

Deutsche Forschungsgemeinschaft (SPP 1927)

  • Roland Lill

European Commission (iNEXT 653706)

  • Lucia Banci

Deutsche Forschungsgemeinschaft (SFB 987)

  • Ulrich Mühlenhoff
  • Roland Lill

European strategy forum on research infrastructures (Instruct)

  • Lucia Banci

LOEWE program of state Hesse, Germany (Synmikro)

  • Roland Lill

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Nikolaus Pfanner, University of Freiburg, Germany

Publication history

  1. Received: April 6, 2016
  2. Accepted: August 8, 2016
  3. Accepted Manuscript published: August 17, 2016 (version 1)
  4. Version of Record published: September 7, 2016 (version 2)
  5. Version of Record updated: September 8, 2016 (version 3)

Copyright

© 2016, Uzarska et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,069
    Page views
  • 571
    Downloads
  • 82
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Marta A Uzarska
  2. Veronica Nasta
  3. Benjamin D Weiler
  4. Farah Spantgar
  5. Simone Ciofi-Baffoni
  6. Maria Rosaria Saviello
  7. Leonardo Gonnelli
  8. Ulrich Mühlenhoff
  9. Lucia Banci
  10. Roland Lill
(2016)
Mitochondrial Bol1 and Bol3 function as assembly factors for specific iron-sulfur proteins
eLife 5:e16673.
https://doi.org/10.7554/eLife.16673
  1. Further reading

Further reading

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Edmundo G Vides, Ayan Adhikari ... Suzanne R Pfeffer
    Research Advance

    Activating mutations in the Leucine Rich Repeat Kinase 2 (LRRK2) cause Parkinson's disease and previously we showed that activated LRRK2 phosphorylates a subset of Rab GTPases (Steger et al., 2017). Moreover, Golgi-associated Rab29 can recruit LRRK2 to the surface of the Golgi and activate it there for both auto- and Rab substrate phosphorylation. Here we define the precise Rab29 binding region of the LRRK2 Armadillo domain between residues 360-450 and show that this domain, termed 'Site #1', can also bind additional LRRK2 substrates, Rab8A and Rab10. Moreover, we identify a distinct, N-terminal, higher affinity interaction interface between LRRK2 phosphorylated Rab8 and Rab10 termed 'Site #2', that can retain LRRK2 on membranes in cells to catalyze multiple, subsequent phosphorylation events. Kinase inhibitor washout experiments demonstrate that rapid recovery of kinase activity in cells depends on the ability of LRRK2 to associate with phosphorylated Rab proteins, and phosphorylated Rab8A stimulates LRRK2 phosphorylation of Rab10 in vitro. Reconstitution of purified LRRK2 recruitment onto planar lipid bilayers decorated with Rab10 protein demonstrates cooperative association of only active LRRK2 with phospho-Rab10-containing membrane surfaces. These experiments reveal a feed-forward pathway that provides spatial control and membrane activation of LRRK2 kinase activity.

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Andrea Volante, Juan Carlos Alonso, Kiyoshi Mizuuchi
    Research Article Updated

    Three-component ParABS partition systems ensure stable inheritance of many bacterial chromosomes and low-copy-number plasmids. ParA localizes to the nucleoid through its ATP-dependent nonspecific DNA-binding activity, whereas centromere-like parS-DNA and ParB form partition complexes that activate ParA-ATPase to drive the system dynamics. The essential parS sequence arrangements vary among ParABS systems, reflecting the architectural diversity of their partition complexes. Here, we focus on the pSM19035 plasmid partition system that uses a ParBpSM of the ribbon-helix-helix (RHH) family. We show that parSpSM with four or more contiguous ParBpSM-binding sequence repeats is required to assemble a stable ParApSM-ParBpSM complex and efficiently activate the ParApSM-ATPase, stimulating complex disassembly. Disruption of the contiguity of the parSpSM sequence array destabilizes the ParApSM-ParBpSM complex and prevents efficient ATPase activation. Our findings reveal the unique architecture of the pSM19035 partition complex and how it interacts with nucleoid-bound ParApSM-ATP.