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Characterizing nucleic acid association with bacterial membrane vesicles and their transfer to host cells
Author(s) -
Rodriguez Blanca Victoria,
Kuehn Meta J.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.669.15
Subject(s) - rna , nucleic acid , biology , microbiology and biotechnology , vesicle , extracellular vesicle , bacteria , microvesicles , small rna , pathogen , microrna , cell , biochemistry , gene , genetics , membrane
Numerous studies have shown that extracellular nucleic acids can be secreted by bacteria via membrane vesicles (MVs) which can serve as protected vehicles for nucleic acid transfer into host cells. MVs are known to carry biologically active macromolecules that play a significant role in bacteria‐host cell communication. A potential role for prokaryotic MV‐associated RNA in pathogen‐host interactions was proposed recently but intensive investigation into this hypothesis has not been conducted. It has been revealed that vesicles contain a distinct profile of RNA molecules relative to the parent cell RNA profile of multiple bacterial organisms, thereby suggesting a selective export or protective mechanism for RNAs associated with vesicles. Previous studies have highlighted the immunomodulatory characteristics of RNA produced by the human pathogen Staphylococcus aureus during infection of host immune cells but it is currently unknown whether immunomodulatory RNAs are secreted in association with MVs. As such, we are interested in gaining a thorough understanding of the identity, selection, localization, and host cell interaction of MV‐associated RNAs derived from S. aureus . Using electrophoretic analysis and microscopy imaging approaches, I have found that RNAs are associated with purified MVs produced by S. aureus . By size distribution analysis, I discovered that the MV‐associated RNAs appeared to be <50 nucleotides in length, consistent with previously published reports for MVs produced by other bacteria. When examining the functional roles of MV associated RNA, I discovered that S. aureus MVs induced the production of a cytokine that is typically induced in response to signaling cascades initiated by nucleic acid receptors in cultured macrophage cells. Thus, my results suggest a role for MV‐associated nucleic acids in stimulating a host immune response that could be distal to the site of infection. Based on these findings, I hypothesize that MVs are enriched with nuclease‐protected nucleic acid fragments containing immunostimulatory sequences. Using RNA‐seq and qRT‐PCR of MV RNA cargo, those sequences can be identified. Nuclease protection assays suggest a portion of the nucleic acids associated with the MVs are protected by a location inside the vesicle membrane or by association with vesicle cargo proteins. By screening for the presence and importance of RNA‐binding proteins in the MVs, I can identify mechanisms of RNA selection and protection. This research will uncover mechanisms of RNA association with bacterial MVs and mechanisms of host immune cell modulation by the MV‐associated RNAs. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .