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Updated: Jun 29, 2025

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages
Published on: November 3, 2014
Lipopolysaccharide Regulates the Macrophage RNA-Binding Proteome
Deepali Rathore1, Matthew J Marino1, Jiraphorn Issara-Amphorn1
1Functional Cellular Networks Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, United States.
This study reveals how RNA-protein interactions regulate innate immunity. Researchers identified key RNA binding proteins (RBPs) that change their RNA binding activity after lipopolysaccharide (LPS) stimulation in macrophages.
Area of Science:
- Molecular Biology
- Immunology
- Proteomics
Background:
- RNA-protein interactions modulate cellular signaling pathways.
- Specific RNA-protein interactions regulate post-transcriptional control during innate immune responses.
Purpose of the Study:
- Investigate changes in the RNA-bound proteome of mouse macrophages after lipopolysaccharide (LPS) treatment.
- Identify RNA binding proteins (RBPs) involved in innate immune signaling.
Main Methods:
- Utilized Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) for quantitative proteomics.
- Employed RNA sequencing, Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS), and RNP complex purification.
- Analyzed RBP-RNA binding sites at amino acid resolution using Nuclease P1 digestion.
Main Results:
- Identified significantly affected RBPs in macrophages post-LPS stimulation.
- Correlated changes in transcript and protein abundance with altered protein-RNA binding.
- Highlighted Il1α, MARCKS, and ACOD1 as potential RBPs in innate immune signaling.
Conclusions:
- RNA-protein interactions play a crucial role in regulating innate immune signaling.
- The study provides a framework for investigating RBP-RNA interactions in immunity.
- Identified specific RBP candidates for further functional studies in innate immunity.
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