Purification, identification, and functional analysis of polysomes from the human pathogen Staphylococcus aureus

Régine Brielle 1 Marie-Laure Pinel-Marie 1, * Sophie Chat 2 Reynald Gillet 3, 2 Brice Felden 1, *
* Auteur correspondant
1 Fonction, structure et inactivation d'ARN bactériens
2 IGDR - Institut de Génétique et Développement de Rennes
3 IUF - Institut Universitaire de France
Abstract : Polysomes are macromolecular complexes made up of multiple ribosomes simultaneously translating a single mRNA into polypeptide chains. Together, the cellular mRNAs translated in this way are referred to 'translatome.' Translation determines a cell's overall gene expression profile. Studying translatome leads to a better understanding of the translational machinery and of its complex regulatory pathways. Given its fundamental role in cell homeostasis and division, bacterial translation is an important target for antibiotics. However, there are no detailed protocols for polysome purification from Staphylococcus aureus, the human pathogen responsible for the majority of multi-drug resistance issues. We therefore developed methods for the isolation of active polysomes, ribosomes, and ribosomal subunits, examining the purity and quality of each fraction and monitoring polysomal activity during protein synthesis. These steps are mandatory for the use of purified S. aureus polysomes and ribosomes for structural studies or for genome-scale analysis of most translated mRNAs.
Keywords : Polysome Ribosome Staphylococcus aureus Sucrose gradient purification translation Translatome
Type de document :
Article dans une revue
Methods, Elsevier, 2017, 117, pp.59-66. 〈10.1016/j.ymeth.2016.10.003〉
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https://hal-univ-rennes1.archives-ouvertes.fr/hal-01502132
Contributeur : Laurent Jonchère <>
Soumis le : jeudi 21 septembre 2017 - 15:28:52
Dernière modification le : mercredi 16 mai 2018 - 11:23:52

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UNIV-RENNES1 | IGDR | U835 | BIOSIT | UR1-UFR-SVE | UNIV-RENNES

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Régine Brielle, Marie-Laure Pinel-Marie, Sophie Chat, Reynald Gillet, Brice Felden. Purification, identification, and functional analysis of polysomes from the human pathogen Staphylococcus aureus. Methods, Elsevier, 2017, 117, pp.59-66. 〈10.1016/j.ymeth.2016.10.003〉. 〈hal-01502132〉

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