A concerted mechanism involving ACAT and SREBPs by which oxysterols deplete accessible cholesterol to restrict microbial infection

Abstract

Most of the cholesterol in the plasma membranes (PMs) of animal cells is sequestered through interactions with phospholipids and transmembrane domains of proteins. However, as cholesterol concentration rises above the PM's sequestration capacity, a new pool of cholesterol, called accessible cholesterol, emerges. The transport of accessible cholesterol between the PM and the endoplasmic reticulum (ER) is critical to maintain cholesterol homeostasis. This pathway has also been implicated in the suppression of both bacterial and viral pathogens by immunomodulatory oxysterols. Here, we describe a mechanism of depletion of accessible cholesterol from PMs by the oxysterol 25-hydroxycholesterol (25HC). We show that 25HC-mediated activation of acyl coenzyme A: cholesterol acyltransferase (ACAT) in the ER creates an imbalance in the equilibrium distribution of accessible cholesterol between the ER and PM. This imbalance triggers the rapid internalization of accessible cholesterol from the PM, which is sustained for long periods of time through 25HC-mediated suppression of SREBPs and continued activation of ACAT. In support of a physiological role for this mechanism, 25HC failed to suppress Zika virus and human coronavirus infection in ACAT-deficient cells, and Listeria monocytogenes infection in ACAT-deficient cells and mice. We propose that selective depletion of accessible PM cholesterol triggered by ACAT activation and sustained through SREBP suppression underpins the immunological activities of 25HC and a functionally related class of oxysterols.

Data availability

All reagents generated in this study are available from the authors with a completed Materials Transfer Agreement. No datasets were generated in this study that required deposition in data repositories. Original, uncropped scans of all immunoblots shown in this study are included in the Source Data Files attached to the respective Figures. The raw data (including replicates and statistics) for all graphs shown in this study are included in the Source Data Files attached to the respective Figures

Article and author information

Author details

  1. David B Heisler

    Department of Microbiology, The 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:" 0000-0002-6482-4215
  2. Kristen A Johnson

    Department of Molecular Genetics, The 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:" 0000-0003-2635-7406
  3. Duo H Ma

    Department of Microbiology, The 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:" 0000-0002-6736-551X
  4. Maikke B Ohlson

    Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  5. Lishu Zhang

    Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  6. Michelle Tran

    Department of Molecular Genetics, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  7. Chase D Corley

    Department of Molecular Genetics, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  8. Michael E Abrams

    Department of Micr, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  9. Jeffrey G McDonald

    Department of Molecular Genetics, The University of Texas Southwestern Medical Center, Dallas, United States
    Competing interests
    No competing interests declared.
  10. 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
  11. Neal M Alto

    Department of Microbiology, The University of Texas Southwestern Medical Center, Dallas, United States
    For correspondence
    neal.alto@utsouthwestern.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7602-3853
  12. Arun Radhakrishnan

    Department of Molecular Genetics, The University of Texas Southwestern Medical Center, Dallas, United States
    For correspondence
    arun.radhakrishnan@utsouthwestern.edu
    Competing interests
    Arun Radhakrishnan, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7266-7336

Funding

National Institutes of Health (AI158357)

  • Neal M Alto
  • Arun Radhakrishnan

National Institutes of Health (HL160487)

  • Jeffrey G McDonald
  • Arun Radhakrishnan

National Institutes of Health (AI083359)

  • Neal M Alto

National Institutes of Health (AI158124)

  • John W Schoggins

National Institutes of Health (5T32AI007520)

  • David B Heisler

Welch Foundation (I-1731)

  • Neal M Alto

Welch Foundation (I-1793)

  • Arun Radhakrishnan

Fondation Leducq (19CVD04)

  • Arun Radhakrishnan

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

Ethics

Animal experimentation: All animal experiments were performed with the approval of the Institutional Animal Care & Use Committee (IACUC) at the University of Texas Southwestern Medical Center (Approval Reference Number: APN102346).

Reviewing Editor

  1. Suzanne R Pfeffer, Stanford University, United States

Publication history

  1. Received: September 17, 2022
  2. Accepted: January 25, 2023
  3. Accepted Manuscript published: January 25, 2023 (version 1)

Copyright

© 2023, Heisler 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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  1. David B Heisler
  2. Kristen A Johnson
  3. Duo H Ma
  4. Maikke B Ohlson
  5. Lishu Zhang
  6. Michelle Tran
  7. Chase D Corley
  8. Michael E Abrams
  9. Jeffrey G McDonald
  10. John W Schoggins
  11. Neal M Alto
  12. Arun Radhakrishnan
(2023)
A concerted mechanism involving ACAT and SREBPs by which oxysterols deplete accessible cholesterol to restrict microbial infection
eLife 12:e83534.
https://doi.org/10.7554/eLife.83534
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