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Environmental Plasticity Of Staphylococcus aureus Extracellular Vesicles RNA Content

Abstract : Introduction: Bacterial extracellular vesicles (EVs) carry various macromolecules able to affect host-pathogen interactions, such as RNAs. Staphylococcus aureus, an important human and animal pathogen, releases EVs whose RNA content is still unkown. Here, we adress what classes of RNAs compose S. aureus EVs. Methods: S. aureus strain HG003 was cultured in Brain Heart Infusion medium under different in vitro conditions: early- and late-stationary phases, in the presence or absence of a sublethal concentration of vancomycin (0.5 μg/mL). EVswere purified from cell-free culture supernatants using density gradient ultracentrifugation. Bacterial and EV samples were submitted to phenolchloroform RNA extraction, DNAse treatment, and library preparation (Ovation Prokaryotic RNA-Seq, Nugen, rRNA depletion). Sequencing was performed using Illumina, NextSeq500, 75 cycles, single reading, High Output. Results: Particle yields were similar between conditions, however, EVs from late-stationary phases were ∼55% ,larger. On average, 78.0% of HG003 annotated genes were identified in EVs, while only ∼5% presented - 90% read coverage. Highly covered EV RNAs included mRNAs coding for virulence-factors (hld, agrBCD, psmB1, sbi, spa, isaB), ribosomal proteins, transcriptional regulators, and metabolic enzymes. sRNAs were also detected, including the bona fide rsaC. Interestingly, several of these RNAs were shown to belong to the same transcriptional units in S. aureus. Both nature and abundance of the RNAs in EVs were dramatically affected by growth conditions, whereas much less in the parent cells. Finally, the RNA abundance pattern differed between EVs and parent cells. Summary/Conclusion: To our knowledge, this is the first work characterizing the RNA cargo of S. aureus EVs. Our findings show that EV RNAs are shaped by the environment, and suggest the selective packaging of RNAs into EVs. Finally, this study also shedds light to the possible roles of potentially functional RNAs in S. aureus EVs, notably in host-pathogen interactions.
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https://hal.inrae.fr/hal-03259481
Contributor : Anne Giboulot Connect in order to contact the contributor
Submitted on : Monday, June 14, 2021 - 11:26:13 AM
Last modification on : Monday, August 16, 2021 - 5:34:01 PM

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Distributed under a Creative Commons Attribution - NonCommercial - NoDerivatives 4.0 International License

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  • HAL Id : hal-03259481, version 1

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Brenda Silva Rosa da Luz, Aurélie Nicolas, Svetlana Chabelskaya, Vinicius de Rezende Rodovalho, Yves Le Loir, et al.. Environmental Plasticity Of Staphylococcus aureus Extracellular Vesicles RNA Content. ISEV2021 Annual Meeting, The International Society For Extracellular Vesicles (ISEV), May 2021, Virtual United States, United States. ⟨hal-03259481⟩

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