Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in .
Fiche publication
Date publication
janvier 2021
Journal
Frontiers in molecular biosciences
Auteurs
Membres identifiés du Cancéropôle Est :
Dr ROMBY Pascale, Dr YUSUPOV Marat
Tous les auteurs :
Belinite M, Khusainov I, Soufari H, Marzi S, Romby P, Yusupov M, Hashem Y
Lien Pubmed
Résumé
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 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 preserving favorable conformation of the helix 44.
Mots clés
RNA stability, Staphylococcus aureus, ribosomal RNA, ribosome 70 S, translation
Référence
Front Mol Biosci. 2021 ;8:738752