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Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus .
- Source :
-
Frontiers in molecular biosciences [Front Mol Biosci] 2021 Nov 18; Vol. 8, pp. 738752. Date of Electronic Publication: 2021 Nov 18 (Print Publication: 2021). - Publication Year :
- 2021
-
Abstract
- Cryo-electron microscopy is now used as a method of choice in structural biology for studying protein synthesis, a process mediated by the ribosome machinery. In order to achieve high-resolution structures using this approach, one needs to obtain homogeneous and stable samples, which requires optimization of ribosome purification in a species-dependent manner. This is especially critical for the bacterial small ribosomal subunit that tends to be unstable in the absence of ligands. Here, we report a protocol for purification of stable 30 S from the Gram-positive bacterium Staphylococcus aureus and its cryo-EM structures: in presence of spermidine at a resolution ranging between 3.4 and 3.6 Å and in its absence at 5.3 Å. Using biochemical characterization and cryo-EM, we demonstrate the importance of spermidine for stabilization of the 30 S via preserving favorable conformation of the helix 44.<br />Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2021 Belinite, Khusainov, Soufari, Marzi, Romby, Yusupov and Hashem.)
Details
- Language :
- English
- ISSN :
- 2296-889X
- Volume :
- 8
- Database :
- MEDLINE
- Journal :
- Frontiers in molecular biosciences
- Publication Type :
- Academic Journal
- Accession number :
- 34869582
- Full Text :
- https://doi.org/10.3389/fmolb.2021.738752