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

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.