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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eLife Sciences Publications Ltd
2024-12-01
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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. |
| format | Article |
| id | doaj-art-aa7c027de3ef42aeb85ccf279c8490fa |
| institution | OA Journals |
| issn | 2050-084X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| 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 |