Formin-2 drives polymerisation of actin filaments enabling segregation of apicoplasts and cytokinesis in Plasmodium falciparum

  1. Johannes Felix Stortz
  2. Mario Del Rosario
  3. Mirko Singer
  4. Jonathan M Wilkes
  5. Markus Meissner  Is a corresponding author
  6. Sujaan Das  Is a corresponding author
  1. University of Glasgow, United Kingdom
  2. Ludwig-Maximillians University, Germany

Abstract

Plasmodium falciparum actin, apart from its role in erythrocyte invasion, is implicated in endocytosis, cytokinesis and inheritance of the chloroplast-like organelle - the apicoplast. However, the inability to visualise filamentous actin (F-actin) dynamics, a limitation we recently overcame for Toxoplasma (Periz et al, 2017), restricted characterisation of both F-actin and actin regulatory proteins. Here, we expressed and validated actin-binding chromobodies as F-actin-sensors in Plasmodium falciparum and characterised in-vivo actin dynamics. F-actin could be chemically modulated, and genetically disrupted upon conditionally deleting actin-1. In a comparative approach, we demonstrate that Formin-2, a predicted nucleator of F-actin, is responsible for apicoplast inheritance in both Plasmodium and Toxoplasma, and additionally mediates efficient cytokinesis in Plasmodium. Finally, time-averaged local intensity measurements of F-actin in Toxoplasma conditional mutants revealed molecular determinants of spatiotemporally regulated F-actin flow. Together, our data indicate that Formin-2 is the primary F-actin nucleator during apicomplexan intracellular growth, mediating multiple essential functions.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Proteomes of interest (Supplementary File 2) were downloaded from the UniProt-KB website (www.uniprot.org).

Article and author information

Author details

  1. Johannes Felix Stortz

    Wellcome Centre for Integrative Parasitology, University of Glasgow, Glasgow, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Mario Del Rosario

    Division of Infection and Immunity, Institute of Biomedical Life Sciences, University of Glasgow, Glasgow, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0430-1463
  3. Mirko Singer

    Faculty of Veterinary Medicine, Ludwig-Maximillians University, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Jonathan M Wilkes

    Wellcome Centre for Molecular Parasitology, University of Glasgow, Glasgow, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  5. Markus Meissner

    Faculty of Veterinary Medicine, Ludwig-Maximillians University, Munich, Germany
    For correspondence
    Markus.Meissner@para.vetmed.uni-muenchen.de
    Competing interests
    The authors declare that no competing interests exist.
  6. Sujaan Das

    Faculty of Veterinary Medicine, Ludwig-Maximillians University, Munich, Germany
    For correspondence
    Sujaan.Das@para.vetmed.uni-muenchen.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6466-4258

Funding

H2020 Excellent Science (ERC-2012-StG 309255-EndoTox)

  • Markus Meissner

Wellcome (Wellcome Senior Fellowship 103875/Z/14/Z)

  • Markus Meissner

Horizon 2020 Framework Programme (LMU Fellowship H2020-MSCA-COFUND-2016-754388)

  • Sujaan Das

Wellcome (085349 - Core funding for the WCMP)

  • Johannes Felix Stortz
  • Mario Del Rosario
  • Jonathan M Wilkes
  • Markus Meissner
  • Sujaan Das

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

Reviewing Editor

  1. Anna Akhmanova, Utrecht University, Netherlands

Version history

  1. Received: June 5, 2019
  2. Accepted: July 17, 2019
  3. Accepted Manuscript published: July 19, 2019 (version 1)
  4. Version of Record published: August 9, 2019 (version 2)

Copyright

© 2019, Felix Stortz 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

  • 3,011
    views
  • 389
    downloads
  • 26
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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. Johannes Felix Stortz
  2. Mario Del Rosario
  3. Mirko Singer
  4. Jonathan M Wilkes
  5. Markus Meissner
  6. Sujaan Das
(2019)
Formin-2 drives polymerisation of actin filaments enabling segregation of apicoplasts and cytokinesis in Plasmodium falciparum
eLife 8:e49030.
https://doi.org/10.7554/eLife.49030

Share this article

https://doi.org/10.7554/eLife.49030

Further reading

    1. Microbiology and Infectious Disease
    Edited by Olivier Silvie et al.
    Collection

    eLife has recently published a wide range of papers on malaria, covering a diversity of themes including parasite biology, epidemiology, immunology, drugs and vaccines.

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Natalia Dolgova, Eva-Maria E Uhlemann ... Oleg Y Dmitriev
    Research Article

    Mediator of ERBB2-driven Cell Motility 1 (MEMO1) is an evolutionary conserved protein implicated in many biological processes; however, its primary molecular function remains unknown. Importantly, MEMO1 is overexpressed in many types of cancer and was shown to modulate breast cancer metastasis through altered cell motility. To better understand the function of MEMO1 in cancer cells, we analyzed genetic interactions of MEMO1 using gene essentiality data from 1028 cancer cell lines and found multiple iron-related genes exhibiting genetic relationships with MEMO1. We experimentally confirmed several interactions between MEMO1 and iron-related proteins in living cells, most notably, transferrin receptor 2 (TFR2), mitoferrin-2 (SLC25A28), and the global iron response regulator IRP1 (ACO1). These interactions indicate that cells with high MEMO1 expression levels are hypersensitive to the disruptions in iron distribution. Our data also indicate that MEMO1 is involved in ferroptosis and is linked to iron supply to mitochondria. We have found that purified MEMO1 binds iron with high affinity under redox conditions mimicking intracellular environment and solved MEMO1 structures in complex with iron and copper. Our work reveals that the iron coordination mode in MEMO1 is very similar to that of iron-containing extradiol dioxygenases, which also display a similar structural fold. We conclude that MEMO1 is an iron-binding protein that modulates iron homeostasis in cancer cells.