Rapid structural analysis of bacterial ribosomes in situ
Abstract Rapid structural analysis of purified proteins and their complexes has become increasingly common thanks to key methodological advances in cryo-electron microscopy (cryo-EM) and associated data processing software packages. In contrast, analogous structural analysis in cells via cryo-electr...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-07586-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832571386103595008 |
|---|---|
| author | Barrett M. Powell Tyler S. Brant Joseph H. Davis Shyamal Mosalaganti |
| author_facet | Barrett M. Powell Tyler S. Brant Joseph H. Davis Shyamal Mosalaganti |
| author_sort | Barrett M. Powell |
| collection | DOAJ |
| description | Abstract Rapid structural analysis of purified proteins and their complexes has become increasingly common thanks to key methodological advances in cryo-electron microscopy (cryo-EM) and associated data processing software packages. In contrast, analogous structural analysis in cells via cryo-electron tomography (cryo-ET) remains challenging due to critical technical bottlenecks, including low-throughput sample preparation and imaging, and laborious data processing methods. Here, we describe a rapid in situ cryo-ET sample preparation and data analysis workflow that results in the routine determination of sub-nm resolution ribosomal structures. We apply this workflow to E. coli, producing a 5.8 Å structure of the 70S ribosome from cells in less than 10 days and facilitating the discovery of a minor population of 100S-like disomes. We envision our approach to be widely applicable to related bacterial samples. |
| format | Article |
| id | doaj-art-805cfbb40f5b44ae83047fec59e442a4 |
| institution | Kabale University |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-805cfbb40f5b44ae83047fec59e442a42025-02-02T12:37:21ZengNature PortfolioCommunications Biology2399-36422025-01-01811910.1038/s42003-025-07586-yRapid structural analysis of bacterial ribosomes in situBarrett M. Powell0Tyler S. Brant1Joseph H. Davis2Shyamal Mosalaganti3Department of Biology, Massachusetts Institute of TechnologyLife Sciences Institute, University of MichiganDepartment of Biology, Massachusetts Institute of TechnologyLife Sciences Institute, University of MichiganAbstract Rapid structural analysis of purified proteins and their complexes has become increasingly common thanks to key methodological advances in cryo-electron microscopy (cryo-EM) and associated data processing software packages. In contrast, analogous structural analysis in cells via cryo-electron tomography (cryo-ET) remains challenging due to critical technical bottlenecks, including low-throughput sample preparation and imaging, and laborious data processing methods. Here, we describe a rapid in situ cryo-ET sample preparation and data analysis workflow that results in the routine determination of sub-nm resolution ribosomal structures. We apply this workflow to E. coli, producing a 5.8 Å structure of the 70S ribosome from cells in less than 10 days and facilitating the discovery of a minor population of 100S-like disomes. We envision our approach to be widely applicable to related bacterial samples.https://doi.org/10.1038/s42003-025-07586-y |
| spellingShingle | Barrett M. Powell Tyler S. Brant Joseph H. Davis Shyamal Mosalaganti Rapid structural analysis of bacterial ribosomes in situ Communications Biology |
| title | Rapid structural analysis of bacterial ribosomes in situ |
| title_full | Rapid structural analysis of bacterial ribosomes in situ |
| title_fullStr | Rapid structural analysis of bacterial ribosomes in situ |
| title_full_unstemmed | Rapid structural analysis of bacterial ribosomes in situ |
| title_short | Rapid structural analysis of bacterial ribosomes in situ |
| title_sort | rapid structural analysis of bacterial ribosomes in situ |
| url | https://doi.org/10.1038/s42003-025-07586-y |
| work_keys_str_mv | AT barrettmpowell rapidstructuralanalysisofbacterialribosomesinsitu AT tylersbrant rapidstructuralanalysisofbacterialribosomesinsitu AT josephhdavis rapidstructuralanalysisofbacterialribosomesinsitu AT shyamalmosalaganti rapidstructuralanalysisofbacterialribosomesinsitu |