The Human RNA-Binding Proteome and Its Dynamics during Translational Arrest

Area of Science:

• Molecular Biology
• Biochemistry
• Genomics

Background:

• Protein-RNA interactions are crucial for numerous biological processes.
• Existing methods lack universality for purifying protein-RNA complexes.
• Understanding these interactions is key to deciphering cellular functions.

Purpose of the Study:

• To introduce XRNAX, a novel method for generic purification of protein-crosslinked RNA.
• To demonstrate the versatility of XRNAX for studying protein-RNA interactions.
• To characterize the composition and dynamics of RNA-bound proteomes.

Main Methods:

• Development of the XRNAX (crosslinked RNA purification) method.
• Application of transcriptomic and proteomic approaches to analyze purified complexes.
• Integration of XRNAX with crosslinking immunoprecipitation sequencing (CLIP-seq).

Main Results:

• XRNAX successfully captures all RNA biotypes.
• Characterization of sub-proteomes interacting with coding and non-coding RNAs (ncRNAs).
• Identification of hundreds of protein-RNA interfaces and dynamic remodeling of the RNA-bound proteome under stress.
• Validation of ncRNA interactions with lamin B1 and EXOSC2 using XRNAX-CLIP-seq.

Conclusions:

• XRNAX provides a versatile and resourceful approach for studying protein-RNA interactions.
• The method facilitates a deeper understanding of the structural and compositional aspects of RNA-biology.
• XRNAX enables the study of dynamic changes in RNA-protein complexes.