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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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Bibliographic Details
Main Authors: Liu, Jianfang (Author) , McRae, Ewan K. S. (Author) , Zhang, Meng (Author) , Geary, Cody (Author) , Andersen, Ebbe Sloth (Author) , Ren, Gang (Author)
Format: Article (Journal)
Language:English
Published: 21 October 2024
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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Summary: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.
Item Description:Gesehen am 19.08.2025
Physical Description:Online Resource
ISSN:2041-1723
DOI:10.1038/s41467-024-52914-1

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