A quantitative framework for structural interpretation of DMS reactivity.

Dimethyl sulfate (DMS) chemical mapping is widely used for probing RNA structure, with low reactivity interpreted as Watson-Crick (WC) base pairs and high reactivity as unpaired nucleotides. Despite its widespread use, a quantitative understanding of how DMS reactivity relates to specific RNA 3D structural features remains incomplete. To address this gap, we systematically analyzed DMS reactivity patterns with a massive library of 7,500 RNA constructs containing two-way junctions with known 3D structures.