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Defining STING-sterol interactions with chemoproteomics.

Ian Ford1,2, Miranda Villanueva3,2, Min Sub Lee1,4

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Cellular cholesterol metabolism regulates the Stimulator of interferon genes (STING) pathway. Sterol synthesis impacts STING activity and localization, revealing new therapeutic targets for autoinflammatory diseases.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Cellular Metabolism

Background:

  • Stimulator of interferon genes (STING) is a crucial intracellular receptor for detecting cytosolic DNA and cyclic dinucleotides.
  • STING regulation is vital to prevent excessive type I interferon responses and autoinflammatory conditions.
  • Cholesterol metabolism's role in modulating STING activity is increasingly recognized but not fully understood.

Purpose of the Study:

  • To elucidate the molecular mechanisms linking cholesterol homeostasis and STING activity.
  • To investigate the impact of cholesterol synthesis modulation on STING function.
  • To identify direct interactions between sterols and STING.

Main Methods:

  • Genetic manipulation (gain- and loss-of-function systems) including SCAP-SREBP2 and Srebf2 deletion.
  • Activity-based protein profiling using sterol-mimetic probes.
  • Co-purification assays and subcellular localization studies.

Main Results:

  • Increasing SCAP-SREBP2-mediated cholesterol synthesis had minimal effect on STING activity.
  • Genetic deletion of Srebf2 enhanced basal and ligand-induced type I interferon responses.
  • Direct evidence of STING-sterol binding was observed; VDAC1 was identified as a STING-interacting protein sensitive to sterols. STING localization responded to cellular sterol content changes.

Conclusions:

  • STING can function independently of the SCAP-SREBP2 pathway.
  • Sterol synthesis in the endoplasmic reticulum influences STING activity.
  • Findings support a model where cholesterol recognition amino acid consensus (CARC) motifs mediate cholesterol-dependent STING retention in the ER.