Fiche publication
Date publication
juin 2017
Journal
Nucleic acids research
Auteurs
Membres identifiés du Cancéropôle Est :
Dr ROMBY Pascale
Tous les auteurs :
Tomasini A, Moreau K, Chicher J, Geissmann T, Vandenesch F, Romby P, Marzi S, Caldelari I
Lien Pubmed
Résumé
The virulon of Staphyloccocus aureus is controlled by intricate connections between transcriptional and post-transcriptional regulators including proteins and small non-coding RNAs (sRNAs). Many of the sRNAs regulate gene expression through base-pairings with mRNAs. However, characterization of the direct sRNA targets in Gram-positive bacteria remained a difficult challenge. Here, we have applied and adapted the MS2-affinity purification approach coupled to RNA sequencing (MAPS) to determine the targetome of RsaA sRNA of S. aureus, known to repress the synthesis of the transcriptional regulator MgrA. Several mRNAs were enriched with RsaA expanding its regulatory network. Besides mgrA, several of these mRNAs encode a family of SsaA-like enzymes involved in peptidoglycan metabolism and the secreted anti-inflammatory FLIPr protein. Using a combination of in vivo and in vitro approaches, these mRNAs were validated as direct RsaA targets. Quantitative differential proteomics of wild-type and mutant strains corroborated the MAPS results. Additionally, it revealed that RsaA indirectly activated the synthesis of surface proteins supporting previous data that RsaA stimulated biofilm formation and favoured chronic infections. All together, this study shows that MAPS could also be easily applied in Gram-positive bacteria for identification of sRNA targetome.
Mots clés
Bacterial Proteins, genetics, Base Sequence, Binding Sites, Gene Expression Regulation, Bacterial, Membrane Proteins, genetics, Proteome, genetics, RNA Interference, RNA, Bacterial, RNA, Messenger, RNA, Untranslated, physiology, Staphylococcus aureus, genetics, Transcriptome
Référence
Nucleic Acids Res.. 2017 Jun 20;45(11):6746-6760
