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

Proteins and RNA functionally and physically intersect in multiple biological processes， however， currently no universal method is available to purify protein-RNA complexes. Here， we introduce XRNAX， a method for the generic purification of protein-crosslinked RNA， and demonstrate its versatility to study the composition and dynamics of protein-RNA interactions by various transcriptomic and proteomic approaches. We show that XRNAX captures all RNA biotypes and use this to characterize the sub-proteomes that interact with coding and non-coding RNAs (ncRNAs) and to identify hundreds of protein-RNA interfaces. Exploiting the quantitative nature of XRNAX， we observe drastic remodeling of the RNA-bound proteome during arsenite-induced stress， distinct from autophagy-related changes in the total proteome. In addition， we combine XRNAX with crosslinking immunoprecipitation sequencing (CLIP-seq) to validate the interaction of ncRNA with lamin B1 and EXOSC2. Thus， XRNAX is a resourceful approach to study structural and compositional aspects of protein-RNA interactions to address fundamental questions in RNA-biology.