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Related Experiment Video

Updated: Jan 5, 2026

Monitoring Activation of the Antiviral Pattern Recognition Receptors RIG-I And PKR By Limited Protease Digestion and Native PAGE
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Monitoring Activation of the Antiviral Pattern Recognition Receptors RIG-I And PKR By Limited Protease Digestion and Native PAGE

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RNA binding activates RIG-I by releasing an autorepressed signaling domain.

T H Dickey1, B Song1, A M Pyle1,2

  • 1Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, USA.

Science Advances
|October 17, 2019
PubMed
Summary

Researchers developed a new method to measure retinoic acid-inducible gene I (RIG-I) activation. RNA alone triggers RIG-I signaling, and ATP has a minimal role, offering insights for autoimmune disease therapeutics.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biochemistry

Background:

  • The retinoic acid-inducible gene I (RIG-I) receptor is crucial for innate immunity and a key target for immunotherapies.
  • Current limitations in monitoring RIG-I activation in vitro hinder understanding of its mechanisms and the discovery of related drugs.

Purpose of the Study:

  • To develop a novel method for measuring RIG-I activation in vitro.
  • To investigate the conformational changes associated with RIG-I activation and the role of ligands.

Main Methods:

  • Utilized a fluorescence resonance energy transfer (FRET)-based assay.
  • Employed dual site-specific fluorescent labeling of the RIG-I protein.
  • Measured conformational changes indicating signaling domain release.

Main Results:

  • Demonstrated that RNA alone is sufficient to induce RIG-I signaling domain ejection.
  • Found that this ejection process is reversible.
  • Determined that adenosine triphosphate (ATP) plays a minor role in RIG-I activation.

Conclusions:

  • The developed FRET method allows for in vitro monitoring of RIG-I activation.
  • Findings clarify the role of RNA and ATP in RIG-I signaling.
  • Results provide a basis for understanding RIG-I dysfunction in autoimmune diseases and for designing targeted therapeutics.