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Chemoproteomic Profiling of 8-Oxoguanosine-Sensitive RNA-Protein Interactions

Jennifer Villers1, Eliana McCann Smith1, Amanda N DeLiberto1

  • 1Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.

Biochemistry
|November 27, 2023
PubMed

Insights

Oxidative stress generates 8-oxoguanine (8OG) in RNA, altering RNA-protein interactions. This study identifies proteins that bind to or are repelled by 8OG-modified RNA, revealing impacts on cellular RNA processing.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Cellular nucleic acids are vulnerable to damage from endogenous and exogenous agents.
  • Oxidative stress causes 8-oxoguanine (8OG) accumulation in DNA and RNA.
  • 8OG lesions on mRNA affect translation, but their impact on RNA-protein interactions is not well understood.

Purpose of the Study:

  • To investigate the global effects of 8-oxoguanine (8OG) on RNA-protein binding.
  • To identify specific proteins that interact differently with 8OG-modified RNA compared to unmodified RNA.
  • To characterize the molecular mechanisms underlying these altered interactions.

Main Methods:

  • Application of an RNA chemical proteomics approach.
  • Biochemical and biophysical assays to quantify binding affinities.
  • Analysis of protein binding preferences to 8OG-modified RNA.

Main Results:

  • Identified proteins that preferentially bind to 8OG-modified RNA, including IGF2BP1-3 and hnRNPD.
  • Identified proteins repelled by 8OG modification, such as RBM4.
  • Demonstrated that a single 8OG lesion can abolish RBM4 binding to its target RNA sequence.

Conclusions:

  • Established the molecular consequences of 8OG on cellular RNA-protein binding.
  • Provided a framework for studying the role of RNA oxidation in biological processes.
  • Highlighted the significant impact of RNA oxidation on gene regulation and cellular function.

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