Elucidating the RNA-Protein Interactomes of Target RNAs in Tissue

RNA-protein interactions are key to many aspects of cellular homeostasis and their identification is important to understanding cellular function. Multiple strategies have been developed for the RNA-centric characterization of RNA-protein complexes. However, these studies have all been done in immortalized cell lines that do not capture the complexity of heterogeneous tissue samples. Here, we develop hybridization purification of RNA-protein complexes followed by mass spectrometry (HyPR-MS) for use in tissue samples. We isolated both polyadenylated RNA and the specific long noncoding RNA MALAT1 and characterized their protein interactomes. These results demonstrate the feasibility of HyPR-MS in tissue for the multiplexed characterization of specific RNA-protein complexes.

Automated summary

RNA-protein interactions are vital for cellular homeostasis, and their identification is crucial for understanding cellular function. This study introduces a new method called hybridization purification of RNA-protein complexes followed by mass spectrometry (HyPR-MS), which enables the characterization of specific RNA-protein complexes in tissue samples. The researchers successfully isolated polyadenylated RNA and the long noncoding RNA MALAT1, and characterized their protein interactomes using this technique, demonstrating its feasibility.