Ligand response of guanidine-IV riboswitch at single-molecule level
Riboswitches represent a class of non-coding RNA that possess the unique ability to specifically bind ligands and, in response, regulate gene expression. A recent report unveiled a type of riboswitch, known as the guanidine-IV riboswitch, which responds to guanidine levels to regulate downstream gen...
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eLife Sciences Publications Ltd
2024-12-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/94706 |
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| _version_ | 1850179565895286784 |
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| author | Lingzhi Gao Dian Chen Yu Liu |
| author_facet | Lingzhi Gao Dian Chen Yu Liu |
| author_sort | Lingzhi Gao |
| collection | DOAJ |
| description | Riboswitches represent a class of non-coding RNA that possess the unique ability to specifically bind ligands and, in response, regulate gene expression. A recent report unveiled a type of riboswitch, known as the guanidine-IV riboswitch, which responds to guanidine levels to regulate downstream genetic transcription. However, the precise molecular mechanism through which the riboswitch senses its target ligand and undergoes conformational changes remain elusive. This gap in understanding has impeded the potential applications of this riboswitch. To bridge this knowledge gap, our study investigated the conformational dynamics of the guanidine-IV riboswitch RNA upon ligand binding. We employed single-molecule fluorescence resonance energy transfer (smFRET) to dissect the behaviors of the aptamer, terminator, and full-length riboswitch. Our findings indicated that the aptamer portion exhibited higher sensitivity to guanidine compared to the terminator and full-length constructs. Additionally, we utilized Position-specific Labelling of RNA (PLOR) combined with smFRET to observe, at the single-nucleotide and single-molecule level, the structural transitions experienced by the guanidine-IV riboswitch during transcription. Notably, we discovered that the influence of guanidine on the riboswitch RNA’s conformations was significantly reduced after the transcription of 88 nucleotides. Furthermore, we proposed a folding model for the guanidine-IV riboswitch in the absence and presence of guanidine, thereby providing insights into its ligand-response mechanism. |
| format | Article |
| id | doaj-art-8f71863413f447978dc37a0ac3bfaba5 |
| 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-8f71863413f447978dc37a0ac3bfaba52025-08-20T02:18:28ZengeLife Sciences Publications LtdeLife2050-084X2024-12-011310.7554/eLife.94706Ligand response of guanidine-IV riboswitch at single-molecule levelLingzhi Gao0https://orcid.org/0000-0002-4273-7946Dian Chen1Yu Liu2https://orcid.org/0000-0002-4188-2949State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, ChinaState Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, ChinaState Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, ChinaRiboswitches represent a class of non-coding RNA that possess the unique ability to specifically bind ligands and, in response, regulate gene expression. A recent report unveiled a type of riboswitch, known as the guanidine-IV riboswitch, which responds to guanidine levels to regulate downstream genetic transcription. However, the precise molecular mechanism through which the riboswitch senses its target ligand and undergoes conformational changes remain elusive. This gap in understanding has impeded the potential applications of this riboswitch. To bridge this knowledge gap, our study investigated the conformational dynamics of the guanidine-IV riboswitch RNA upon ligand binding. We employed single-molecule fluorescence resonance energy transfer (smFRET) to dissect the behaviors of the aptamer, terminator, and full-length riboswitch. Our findings indicated that the aptamer portion exhibited higher sensitivity to guanidine compared to the terminator and full-length constructs. Additionally, we utilized Position-specific Labelling of RNA (PLOR) combined with smFRET to observe, at the single-nucleotide and single-molecule level, the structural transitions experienced by the guanidine-IV riboswitch during transcription. Notably, we discovered that the influence of guanidine on the riboswitch RNA’s conformations was significantly reduced after the transcription of 88 nucleotides. Furthermore, we proposed a folding model for the guanidine-IV riboswitch in the absence and presence of guanidine, thereby providing insights into its ligand-response mechanism.https://elifesciences.org/articles/94706RNAriboswitchguanidinesmFRET |
| spellingShingle | Lingzhi Gao Dian Chen Yu Liu Ligand response of guanidine-IV riboswitch at single-molecule level eLife RNA riboswitch guanidine smFRET |
| title | Ligand response of guanidine-IV riboswitch at single-molecule level |
| title_full | Ligand response of guanidine-IV riboswitch at single-molecule level |
| title_fullStr | Ligand response of guanidine-IV riboswitch at single-molecule level |
| title_full_unstemmed | Ligand response of guanidine-IV riboswitch at single-molecule level |
| title_short | Ligand response of guanidine-IV riboswitch at single-molecule level |
| title_sort | ligand response of guanidine iv riboswitch at single molecule level |
| topic | RNA riboswitch guanidine smFRET |
| url | https://elifesciences.org/articles/94706 |
| work_keys_str_mv | AT lingzhigao ligandresponseofguanidineivriboswitchatsinglemoleculelevel AT dianchen ligandresponseofguanidineivriboswitchatsinglemoleculelevel AT yuliu ligandresponseofguanidineivriboswitchatsinglemoleculelevel |