SYSTEMATIC DETECTION OF TERTIARY STRUCTURAL MODULES IN LARGE RNAS AND RNP INTERFACES BY TB-SEQ

Systematic detection of tertiary structural modules in large RNAs and RNP interfaces by Tb-seq

Systematic detection of tertiary structural modules in large RNAs and RNP interfaces by Tb-seq

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Abstract Compact RNA structural motifs control many aspects of gene expression, moen rothbury faucet but we lack methods for finding these structures in the vast expanse of multi-kilobase RNAs.To adopt specific 3-D shapes, many RNA modules must compress their RNA backbones together, bringing negatively charged phosphates into close proximity.This is often accomplished by recruiting multivalent cations (usually Mg2+), which stabilize these sites and neutralize regions of local negative charge.

Coordinated lanthanide ions, such as terbium (III) (Tb3+), can also be recruited to these sites, where they induce efficient RNA cleavage, thereby revealing compact RNA 3-D modules.Until now, Tb3+ cleavage sites were monitored via low-throughput biochemical methods only applicable to small RNAs.Here we present Tb-seq, a high-throughput sequencing method for detecting compact tertiary structures in large RNAs.

Tb-seq detects sharp backbone turns gymnastics wall decals found in RNA tertiary structures and RNP interfaces, providing a way to scan transcriptomes for stable structural modules and potential riboregulatory motifs.

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