Non-averaged single-molecule tertiary structures reveal RNA self-folding through individual-particle cryo-electron tomography
Large-scale and continuous conformational changes in the RNA self-folding process present significant challenges for structural studies, often requiring trade-offs between resolution and observational scope. Here, we utilize individual-particle cryo-electron tomography (IPET) to examine the post-tra...
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| Main Authors: | , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
21 October 2024
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| In: |
Nature Communications
Year: 2024, Volume: 15, Pages: 1-18 |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/s41467-024-52914-1 |
| Online Access: | Verlag, kostenfrei, Volltext: https://doi.org/10.1038/s41467-024-52914-1 Verlag, kostenfrei, Volltext: https://www.nature.com/articles/s41467-024-52914-1 
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| Author Notes: | Jianfang Liu, Ewan K.S. McRae, Meng Zhang, Cody Geary, Ebbe Sloth Andersen & Gang Ren |
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| 520 | |a Large-scale and continuous conformational changes in the RNA self-folding process present significant challenges for structural studies, often requiring trade-offs between resolution and observational scope. Here, we utilize individual-particle cryo-electron tomography (IPET) to examine the post-transcriptional self-folding process of designed RNA origami 6-helix bundle with a clasp helix (6HBC). By avoiding selection, classification, averaging, or chemical fixation and optimizing cryo-ET data acquisition parameters, we reconstruct 120 three-dimensional (3D) density maps from 120 individual particles at an electron dose of no more than 168 e-Å−2, achieving averaged resolutions ranging from 23 to 35 Å, as estimated by Fourier shell correlation (FSC) at 0.5. Each map allows us to identify distinct RNA helices and determine a unique tertiary structure. Statistical analysis of these 120 structures confirms two reported conformations and reveals a range of kinetically trapped, intermediate, and highly compacted states, demonstrating a maturation folding landscape likely driven by helix-helix compaction interactions. | ||
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| 650 | 4 | |a Molecular imaging | |
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| 700 | 1 | |a Ren, Gang |e VerfasserIn |4 aut | |
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