High-throughput determination of RNA tertiary contact thermodynamics by quantitative DMS chemical mapping.

ABSTRACT: Structured RNAs often contain long-range tertiary contacts that are critical to their function. Despite the importance of tertiary contacts, methods to measure their thermodynamics are low throughput or require specialized instruments. Here, we introduce a new quantitative chemical mapping method (qMaPseq) to measure Mg2+-induced formation of tertiary contact thermodynamics in a high-throughput manner using standard biochemistry equipment. With qMaPseq, we measured the ΔG of 98 unique tetraloop/tetraloop receptor (TL/TLR) variants in a one-pot reaction. These results agree well with measurements from specialized instruments (R2= 0.64). Furthermore, the DMS reactivity of the TL directly correlates to the stability of the contact (R2= 0.68), the first direct evidence that a single DMS reacti

SUBMITTER: Lange B 

PROVIDER: S-EPMC11381326 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature