Over-expression screen of interferon-stimulated genes identifies RARRES3 as a restrictor of Toxoplasma gondii infection

  1. Nicholas Rinkenberger
  2. Michael E Abrams
  3. Sumit K Matta
  4. John W Schoggins
  5. Neal M Alto
  6. L David Sibley  Is a corresponding author
  1. Washington University in St. Louis, United States
  2. The University of Texas Southwestern Medical Center, United States
  3. University of Texas Southwestern Medical Center, United States

Abstract

Toxoplasma gondii is an important human pathogen infecting an estimated 1 in 3 people worldwide. The cytokine interferon gamma (IFNγ) is induced during infection and is critical for restricting T. gondii growth in human cells. Growth restriction is presumed to be due to the induction interferon stimulated genes (ISGs) that are upregulated to protect the host from infection. Although there are hundreds of ISGs induced by IFNγ, their individual roles in restricting parasite growth in human cells remain somewhat elusive. To address this deficiency, we screened a library of 414 IFNγ induced ISGs to identify factors that impact T. gondii infection in human cells. In addition to IRF1, which likely acts through induction of numerous downstream genes, we identified RARRES3 as a single factor that restricts T. gondii infection by inducing premature egress of the parasite in multiple human cell lines. Overall, while we successfully identified a novel IFNγ induced factor restricting T. gondii infection, the limited number of ISGs capable of restricting T. gondii infection when individually expressed suggests that IFNγ mediated immunity to T. gondii infection is a complex, multifactorial process.

Data availability

RNASeq data generated here have been deposited to GEO with the accession number GSE181861.

The following data sets were generated

Article and author information

Author details

  1. Nicholas Rinkenberger

    Department of Molecular Microbiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  2. Michael E Abrams

    Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  3. Sumit K Matta

    Department of Molecular Microbiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  4. John W Schoggins

    Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    John W Schoggins, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7944-6800
  5. Neal M Alto

    Microbiology, University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7602-3853
  6. L David Sibley

    Department of Molecular Microbiology, Washington University in St. Louis, St Louis, United States
    For correspondence
    sibley@wustl.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7110-0285

Funding

National Institutes of Health (AI154048)

  • L David Sibley

National Institutes of Health (AI118426)

  • L David Sibley

National Institutes of Health (AI083359)

  • Neal M Alto

Welch Foundation (I-1704)

  • Neal M Alto

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

Reviewing Editor

  1. Russell E Vance, University of California, Berkeley, United States

Publication history

  1. Received: August 17, 2021
  2. Preprint posted: September 5, 2021 (view preprint)
  3. Accepted: December 5, 2021
  4. Accepted Manuscript published: December 6, 2021 (version 1)
  5. Version of Record published: January 25, 2022 (version 2)
  6. Version of Record updated: May 19, 2022 (version 3)

Copyright

© 2021, Rinkenberger 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

  • 880
    Page views
  • 202
    Downloads
  • 3
    Citations

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

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. Nicholas Rinkenberger
  2. Michael E Abrams
  3. Sumit K Matta
  4. John W Schoggins
  5. Neal M Alto
  6. L David Sibley
(2021)
Over-expression screen of interferon-stimulated genes identifies RARRES3 as a restrictor of Toxoplasma gondii infection
eLife 10:e73137.
https://doi.org/10.7554/eLife.73137
  1. Further reading

Further reading

    1. Microbiology and Infectious Disease
    Liheng Yang et al.
    Research Article

    Infections at the maternal-fetal interface can directly harm the fetus and induce complications that adversely impact pregnancy outcomes. Innate immune signaling by both fetal-derived placental trophoblasts and the maternal decidua must provide antimicrobial defenses at this critical interface without compromising its integrity. Here, we developed matched trophoblast and decidua organoids from human placentas to define the relative contributions of these cells to antiviral defenses at the maternal-fetal interface. We demonstrate that trophoblast and decidua organoids basally secrete distinct immunomodulatory factors, including the constitutive release of the antiviral type III interferon IFN-λ2 from trophoblast organoids, and differentially respond to viral infections through the induction of organoid-specific factors. Lastly, we define the differential susceptibility and innate immune signaling of trophoblast and decidua organoids to human cytomegalovirus (HCMV) and develop a co-culture model of trophoblast and decidua organoids which showed that trophoblast-derived factors protect decidual cells from HCMV infection. Our findings establish matched trophoblast and decidua organoids as ex vivo models to study vertically transmitted infections and highlight differences in innate immune signaling by fetal-derived trophoblasts and the maternal decidua.

    1. Cell Biology
    2. Microbiology and Infectious Disease
    Alice L Herneisen et al.
    Research Article

    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.