RNA tertiary structure and conformational dynamics revealed by BASH MaP

The functional effects of an RNA can arise from complex three-dimensional folds known as tertiary structures. However, predicting the tertiary structure of an RNA and whether an RNA adopts distinct tertiary conformations remains challenging. To address this, we developed BASH MaP, a single-molecule...

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Main Authors: Maxim Oleynikov, Samie R Jaffrey
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2024-12-01
Series:eLife
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Online Access:https://elifesciences.org/articles/98540
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author Maxim Oleynikov
Samie R Jaffrey
author_facet Maxim Oleynikov
Samie R Jaffrey
author_sort Maxim Oleynikov
collection DOAJ
description The functional effects of an RNA can arise from complex three-dimensional folds known as tertiary structures. However, predicting the tertiary structure of an RNA and whether an RNA adopts distinct tertiary conformations remains challenging. To address this, we developed BASH MaP, a single-molecule dimethyl sulfate (DMS) footprinting method and DAGGER, a computational pipeline, to identify alternative tertiary structures adopted by different molecules of RNA. BASH MaP utilizes potassium borohydride to reveal the chemical accessibility of the N7 position of guanosine, a key mediator of tertiary structures. We used BASH MaP to identify diverse conformational states and dynamics of RNA G-quadruplexes, an important RNA tertiary motif, in vitro and in cells. BASH MaP and DAGGER analysis of the fluorogenic aptamer Spinach reveals that it adopts alternative tertiary conformations which determine its fluorescence states. BASH MaP thus provides an approach for structural analysis of RNA by revealing previously undetectable tertiary structures.
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spelling doaj-art-aa7c027de3ef42aeb85ccf279c8490fa2025-08-20T02:38:29ZengeLife Sciences Publications LtdeLife2050-084X2024-12-011310.7554/eLife.98540RNA tertiary structure and conformational dynamics revealed by BASH MaPMaxim Oleynikov0https://orcid.org/0000-0002-6892-9743Samie R Jaffrey1https://orcid.org/0000-0003-3615-6958Department of Pharmacology, Weill Medical College, Cornell University, New York, United StatesDepartment of Pharmacology, Weill Medical College, Cornell University, New York, United StatesThe functional effects of an RNA can arise from complex three-dimensional folds known as tertiary structures. However, predicting the tertiary structure of an RNA and whether an RNA adopts distinct tertiary conformations remains challenging. To address this, we developed BASH MaP, a single-molecule dimethyl sulfate (DMS) footprinting method and DAGGER, a computational pipeline, to identify alternative tertiary structures adopted by different molecules of RNA. BASH MaP utilizes potassium borohydride to reveal the chemical accessibility of the N7 position of guanosine, a key mediator of tertiary structures. We used BASH MaP to identify diverse conformational states and dynamics of RNA G-quadruplexes, an important RNA tertiary motif, in vitro and in cells. BASH MaP and DAGGER analysis of the fluorogenic aptamer Spinach reveals that it adopts alternative tertiary conformations which determine its fluorescence states. BASH MaP thus provides an approach for structural analysis of RNA by revealing previously undetectable tertiary structures.https://elifesciences.org/articles/98540DMS probingG-quadruplexchemical probingspinachpolyUGAKT2
spellingShingle Maxim Oleynikov
Samie R Jaffrey
RNA tertiary structure and conformational dynamics revealed by BASH MaP
eLife
DMS probing
G-quadruplex
chemical probing
spinach
polyUG
AKT2
title RNA tertiary structure and conformational dynamics revealed by BASH MaP
title_full RNA tertiary structure and conformational dynamics revealed by BASH MaP
title_fullStr RNA tertiary structure and conformational dynamics revealed by BASH MaP
title_full_unstemmed RNA tertiary structure and conformational dynamics revealed by BASH MaP
title_short RNA tertiary structure and conformational dynamics revealed by BASH MaP
title_sort rna tertiary structure and conformational dynamics revealed by bash map
topic DMS probing
G-quadruplex
chemical probing
spinach
polyUG
AKT2
url https://elifesciences.org/articles/98540
work_keys_str_mv AT maximoleynikov rnatertiarystructureandconformationaldynamicsrevealedbybashmap
AT samierjaffrey rnatertiarystructureandconformationaldynamicsrevealedbybashmap