A method to map changes in bacterial surface composition induced by regulatory RNAs in Escherichia coli and Staphylococcus aureus.
Fiche publication
Date publication
novembre 2014
Journal
Biochimie
Auteurs
Membres identifiés du Cancéropôle Est :
Mr HAMMANN Philippe, Dr ROMBY Pascale
Tous les auteurs :
Hammann P, Parmentier D, Cerciat M, Reimegård J, Helfer AC, Boisset S, Guillier M, Vandenesch F, Wagner EG, Romby P, Fechter P
Lien Pubmed
Résumé
We have adapted a method to map cell surface proteins and to monitor the effect of specific regulatory RNAs on the surface composition of the bacteria. This method involves direct labeling of surface proteins of living bacteria using fluorescent dyes and a subsequent separation of the crude extract by 2D gel electrophoresis. The strategy yields a substantial enrichment in surface proteins over cytoplasmic proteins. We validated this method by monitoring the effect of the regulatory RNA MicA in Escherichia coli, which regulates the synthesis of several outer membrane proteins, and highlighted the role of Staphylococcus aureus RNAIII for the maintenance of cell wall integrity.
Mots clés
Bacterial Outer Membrane Proteins, metabolism, Bacterial Proteins, metabolism, Base Sequence, Carbocyanines, metabolism, Cell Membrane, metabolism, Cell Wall, metabolism, Electrophoresis, Gel, Two-Dimensional, methods, Escherichia coli, genetics, Microscopy, Confocal, Molecular Sequence Data, RNA, Bacterial, genetics, Reproducibility of Results, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Staining and Labeling, methods, Staphylococcus aureus, genetics
Référence
Biochimie. 2014 Nov;106:175-9