Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus

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 optimi...

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Autores principales:	Margarita Belinite, Iskander Khusainov, Heddy Soufari, Stefano Marzi, Pascale Romby, Marat Yusupov, Yaser Hashem
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	ribosome 70 S ribosomal RNA Staphylococcus aureus translation RNA stability Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/23d21fc50ef041a88dc7a44daa03a6f9
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spelling	oai:doaj.org-article:23d21fc50ef041a88dc7a44daa03a6f92021-11-18T07:46:36ZStabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus2296-889X10.3389/fmolb.2021.738752https://doaj.org/article/23d21fc50ef041a88dc7a44daa03a6f92021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmolb.2021.738752/fullhttps://doaj.org/toc/2296-889XCryo-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.Margarita BeliniteMargarita BeliniteMargarita BeliniteIskander KhusainovIskander KhusainovHeddy SoufariStefano MarziPascale RombyMarat YusupovMarat YusupovYaser HashemYaser HashemFrontiers Media S.A.articleribosome 70 Sribosomal RNAStaphylococcus aureustranslationRNA stabilityBiology (General)QH301-705.5ENFrontiers in Molecular Biosciences, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	ribosome 70 S ribosomal RNA Staphylococcus aureus translation RNA stability Biology (General) QH301-705.5
spellingShingle	ribosome 70 S ribosomal RNA Staphylococcus aureus translation RNA stability Biology (General) QH301-705.5 Margarita Belinite Margarita Belinite Margarita Belinite Iskander Khusainov Iskander Khusainov Heddy Soufari Stefano Marzi Pascale Romby Marat Yusupov Marat Yusupov Yaser Hashem Yaser Hashem Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus
description	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.
format	article
author	Margarita Belinite Margarita Belinite Margarita Belinite Iskander Khusainov Iskander Khusainov Heddy Soufari Stefano Marzi Pascale Romby Marat Yusupov Marat Yusupov Yaser Hashem Yaser Hashem
author_facet	Margarita Belinite Margarita Belinite Margarita Belinite Iskander Khusainov Iskander Khusainov Heddy Soufari Stefano Marzi Pascale Romby Marat Yusupov Marat Yusupov Yaser Hashem Yaser Hashem
author_sort	Margarita Belinite
title	Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus
title_short	Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus
title_full	Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus
title_fullStr	Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus
title_full_unstemmed	Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus
title_sort	stabilization of ribosomal rna of the small subunit by spermidine in staphylococcus aureus
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/23d21fc50ef041a88dc7a44daa03a6f9
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Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in Staphylococcus aureus

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