Temporal and thermal profiling of the Toxoplasma proteome implicates parasite Protein Phosphatase 1 in the regulation of Ca2+-responsive pathways

  1. Alice L Herneisen
  2. Zhu-Hong Li
  3. Alex W Chan
  4. Silvia NJ Moreno
  5. Sebastian Lourido  Is a corresponding author
  1. Whitehead Institute for Biomedical Research, United States
  2. University of Georgia, United States

Abstract

Apicomplexan parasites cause persistent mortality and morbidity worldwide through diseases including malaria, toxoplasmosis, and cryptosporidiosis. Ca2+ signaling pathways have been repurposed in these eukaryotic pathogens to regulate parasite-specific cellular processes governing the replicative and lytic phases of the infectious cycle, as well as the transition between them. Despite the presence of conserved Ca2+-responsive proteins, little is known about how specific signaling elements interact to impact pathogenesis. We mapped the Ca2+-responsive proteome of the model apicomplexan T. gondii via time-resolved phosphoproteomics and thermal proteome profiling. The waves of phosphoregulation following PKG activation and stimulated Ca2+ release corroborate known physiological changes but identify specific proteins operating in these pathways. Thermal profiling of parasite extracts identified many expected Ca2+-responsive proteins, such as parasite Ca2+-dependent protein kinases. Our approach also identified numerous Ca2+-responsive proteins that are not predicted to bind Ca2+, yet are critical components of the parasite signaling network. We characterized protein phosphatase 1 (PP1) as a Ca2+-responsive enzyme that relocalized to the parasite apex upon Ca2+ store release. Conditional depletion of PP1 revealed that the phosphatase regulates Ca2+ uptake to promote parasite motility. PP1 may thus be partly responsible for Ca2+-regulated serine/threonine phosphatase activity in apicomplexan parasites.

Data availability

All mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD033765 and 10.6019/PXD033765. All other information is provided in the Supplementary Files.

The following data sets were generated

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Author details

  1. Alice L Herneisen

    Whitehead Institute for Biomedical Research, Cambridge, United States
    Competing interests
    No competing interests declared.
  2. Zhu-Hong Li

    Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, United States
    Competing interests
    No competing interests declared.
  3. Alex W Chan

    Whitehead Institute for Biomedical Research, Cambridge, United States
    Competing interests
    No competing interests declared.
  4. Silvia NJ Moreno

    Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2041-6295
  5. Sebastian Lourido

    Whitehead Institute for Biomedical Research, Cambridge, United States
    For correspondence
    lourido@wi.mit.edu
    Competing interests
    Sebastian Lourido, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5237-1095

Funding

National Institutes of Health (R01AI144369)

  • Sebastian Lourido

National Science Foundation (174530)

  • Alice L Herneisen

National Institutes of Health (R01AI128356)

  • Silvia NJ Moreno

National Institutes of Health (R21AI15493)

  • Silvia NJ Moreno

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

Copyright

© 2022, Herneisen 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.

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https://doi.org/10.7554/eLife.80336

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