Differential analysis of RNA structure probing experiments at nucleotide resolution: uncovering regulatory functions of RNA structure

Abstract

RNAs perform their function by forming specific structures, which can change across cellular conditions. Structure probing experiments combined with next generation sequencing technology have enabled transcriptome-wide analysis of RNA secondary structure in various cellular conditions. Differential analysis of structure probing data in different conditions can reveal the RNA structurally variable regions (SVRs), which is important for understanding RNA functions. Here, we propose DiffScan, a computational framework for normalization and differential analysis of structure probing data in high resolution. DiffScan preprocesses structure probing datasets to remove systematic bias, and then scans the transcripts to identify SVRs and adaptively determines their lengths and locations. The proposed approach is compatible with most structure probing platforms (e.g., icSHAPE, DMS-seq). When evaluated with simulated and benchmark datasets, DiffScan identifies structurally variable regions at nucleotide resolution, with substantial improvement in accuracy compared with existing SVR detection methods. Moreover, the improvement is robust when tested in multiple structure probing platforms. Application of DiffScan in a dataset of multi-subcellular RNA structurome and a subsequent motif enrichment analysis suggest potential links of RNA structural variation and mRNA abundance, possibly mediated by RNA binding proteins such as the serine/arginine rich splicing factors. This work provides an effective tool for differential analysis of RNA secondary structure, reinforcing the power of structure probing experiments in deciphering the dynamic RNA structurome.

DeepFusion: A deep bimodal information fusion network for unraveling protein-RNA interactions using in vivo RNA structures

“…The binding preferences of RBPs depend not only on the RNA sequences but also on the structural features of these RBP binding sites [19] , [20] , [21] , [22] , [23] , [24] , [25] . For example, A G-rich internal loop in the lncRNA Braveheart is critical for binding to a zinc-finger protein CNBP, which is required for the cardiac specification in mice [26] .…”