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Simultaneous mapping of RNA secondary, tertiary, and quaternary structure in living cells by multi-site DMS probing II

ABSTRACT: Identifying tertiary structures and protein binding sites on RNA molecules remains a key challenge in RNA biology. We report a new chemical probing strategy termed multi-site DMS-MaP (msDMS-MaP) that exploits base tautomerization during mutational profiling reverse-transcription to measure typically invisible DMS-induced modifications at the N7 position of G. These N7-G modifications are now resolved concurrently with conventional N1 and N3 modifications with only minor modifications to established protocols, providing a multi-dimensional, single-molecule readout of RNA structure. We show that N7-G reactivity specifically reports on RNA tertiary and quaternary structure, enabling identification of diverse, functionally significant motifs such as cooperatively folding tertiary domains and protein binding sites in living cells. We apply msDMS-MaP to obtain a map of the quaternary structural ensemble of the 7SK non-coding snRNP, revealing unique protein binding sites across three 7SK structural isoforms. msDMS-MaP represents a broadly applicable strategy for enhanced RNA functional motif discovery and characterization.

ORGANISM(S): Homo sapiens

PROVIDER: GSE285334 | GEO | 2026/03/21

REPOSITORIES: GEO

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			Action	DRS
		Other
	GSE285334_HEK293T-7SK-RNP-MaP-Rep1_Hs_RN7SK_profile.txt.gz	Txt
	GSE285334_HEK293T-7SK-RNP-MaP-Rep1_Hs_RN7SK_profile_NT_thresholds.txt.gz	Txt
	GSE285334_HEK293T-7SK-RNP-MaP-Rep1_Hs_RN7SK_profile_RNPMaP_RESULTS.csv.gz	Csv
	GSE285334_HEK293T-7SK-RNP-MaP-Rep2_Hs_RN7SK_profile.txt.gz	Txt

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Simultaneous mapping of RNA secondary, tertiary, and quaternary structure in living cells by multi-site DMS probing I

Project description:Identifying tertiary structures and protein binding sites on RNA molecules remains a key challenge in RNA biology. We report a new chemical probing strategy termed multi-site DMS-MaP (msDMS-MaP) that exploits base tautomerization during mutational profiling reverse-transcription to measure typically invisible DMS-induced modifications at the N7 position of G. These N7-G modifications are now resolved concurrently with conventional N1 and N3 modifications with only minor modifications to established protocols, providing a multi-dimensional, single-molecule readout of RNA structure. We show that N7-G reactivity specifically reports on RNA tertiary and quaternary structure, enabling identification of diverse, functionally significant motifs such as cooperatively folding tertiary domains and protein binding sites in living cells. We apply msDMS-MaP to obtain a map of the quaternary structural ensemble of the 7SK non-coding snRNP, revealing unique protein binding sites across three 7SK structural isoforms. msDMS-MaP represents a broadly applicable strategy for enhanced RNA functional motif discovery and characterization.

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