Mapping RNA-Protein Interactions with Subcellular Resolution Using Colocalization CLIP

RNA. 2024 Apr 24:rna.079890.123. doi: 10.1261/rna.079890.123. Online ahead of print.ABSTRACTRNA binding proteins (RBPs) are essential for RNA metabolism and profoundly impact health and disease. The subcellular organization of RBP interaction networks with target RNAs remains largely unexplored. Here, we develop colocalization CLIP, a method that combines CrossLinking and ImmunoPrecipitation (CLIP) with proximity labeling, to explore in-depth the subcellular RNA interactions of the RNA-binding protein HuR. Using this method, we uncover HuR's dynamic and location-specific interactions with RNA, revealing alterations in sequence preferences and interactions in the nucleus, cytosol, or stress granule compartments. We uncover HuR's unique binding preferences within stress granules during arsenite stress, illuminating intricate interactions that conventional methodologies cannot capture. Overall, coCLIP provides a powerful method for revealing RBP:RNA interactions based on localization and lays the foundation for an advanced understanding of RBP models that incorporate subcellular location as a critical determinant of their functions.PMID:38658162 | DOI:10.1261/rna.079890.123

Source: RNA - April 24, 2024 Category: Genetics & Stem Cells Authors: Soon Yi Shashi S Singh Kathryn Rozen-Gagnon Joseph M Luna Source Type: research