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Article ; Online: Proteomic Profiling Reveals Distinct Bacterial Extracellular Vesicle Subpopulations with Possibly Unique Functionality.

McMillan, Hannah M / Kuehn, Meta J

2022 Volume 89, Issue 1, Page(s) e0168622

Abstract: Bacterial outer membrane vesicles (OMVs) are 20- to 200-nm secreted packages of lipids, small molecules, and proteins that contribute to diverse bacterial processes. In plant systems, OMVs from pathogenic and beneficial strains elicit plant immune ... ...

Abstract	Bacterial outer membrane vesicles (OMVs) are 20- to 200-nm secreted packages of lipids, small molecules, and proteins that contribute to diverse bacterial processes. In plant systems, OMVs from pathogenic and beneficial strains elicit plant immune responses that inhibit seedling growth and protect against future pathogen challenge. Previous studies of OMV-plant interactions suggest functionally important differences in the protein composition of Pseudomonas syringae and Pseudomonas fluorescens OMVs, and that their composition and activity differ as a result of medium culture conditions. Here, we show that plant apoplast-mimicking minimal medium conditions impact OMV protein content dramatically in P. syringae but not in P. fluorescens relative to complete medium conditions. Comparative, 2-way analysis of the four conditions reveals subsets of proteins that may contribute to OMV-mediated bacterial virulence and plant immune activation as well as those involved in bacterial stress tolerance or adaptation to a beneficial relationship with plants. Additional localization enrichment analysis of these subsets suggests the presence of outer-inner membrane vesicles (OIMVs). Collectively, these results reveal distinct differences in bacterial extracellular vesicle cargo and biogenesis routes from pathogenic and beneficial plant bacteria in different medium conditions and point to distinct populations of vesicles with diverse functional roles.
MeSH term(s)	Proteomics/methods ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Bacterial Outer Membrane Proteins/metabolism ; Bacteria/metabolism ; Extracellular Vesicles/metabolism
Chemical Substances	Bacterial Proteins ; Bacterial Outer Membrane Proteins
Language	English
Publishing date	2022-12-19
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	223011-2
ISSN	1098-5336 ; 0099-2240
ISSN (online)	1098-5336
ISSN	0099-2240
DOI	10.1128/aem.01686-22
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Article ; Online: Staphylococcus aureus secretes immunomodulatory RNA and DNA via membrane vesicles.

Rodriguez, Blanca V / Kuehn, Meta J

Scientific reports

2020 Volume 10, Issue 1, Page(s) 18293

Abstract: Bacterial-derived RNA and DNA can function as ligands for intracellular receptor activation and induce downstream signaling to modulate the host response to bacterial infection. The mechanisms underlying the secretion of immunomodulatory RNA and DNA by ... ...

Abstract	Bacterial-derived RNA and DNA can function as ligands for intracellular receptor activation and induce downstream signaling to modulate the host response to bacterial infection. The mechanisms underlying the secretion of immunomodulatory RNA and DNA by pathogens such as Staphylococcus aureus and their delivery to intracellular host cell receptors are not well understood. Recently, extracellular membrane vesicle (MV) production has been proposed as a general secretion mechanism that could facilitate the delivery of functional bacterial nucleic acids into host cells. S. aureus produce membrane-bound, spherical, nano-sized, MVs packaged with a select array of bioactive macromolecules and they have been shown to play important roles in bacterial virulence and in immune modulation through the transmission of biologic signals to host cells. Here we show that S. aureus secretes RNA and DNA molecules that are mostly protected from degradation by their association with MVs. Importantly, we demonstrate that MVs can be delivered into cultured macrophage cells and subsequently stimulate a potent IFN-β response in recipient cells via activation of endosomal Toll-like receptors. These findings advance our understanding of the mechanisms by which bacterial nucleic acids traffic extracellularly to trigger the modulation of host immune responses.
MeSH term(s)	Animals ; DNA, Bacterial/immunology ; Extracellular Vesicles/genetics ; Extracellular Vesicles/immunology ; Interferon-gamma/genetics ; Macrophages/cytology ; Macrophages/immunology ; Macrophages/virology ; Mice ; Particle Size ; RAW 264.7 Cells ; RNA, Bacterial/immunology ; Staphylococcus aureus/genetics ; Staphylococcus aureus/immunology ; Staphylococcus aureus/pathogenicity ; Toll-Like Receptors/genetics ; Virulence
Chemical Substances	DNA, Bacterial ; IFNG protein, mouse ; RNA, Bacterial ; Toll-Like Receptors ; Interferon-gamma (82115-62-6)
Language	English
Publishing date	2020-10-26
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-75108-3
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Article ; Online: Pseudomonas aeruginosa Leucine Aminopeptidase Influences Early Biofilm Composition and Structure via Vesicle-Associated Antibiofilm Activity.

Esoda, Caitlin N / Kuehn, Meta J

mBio

2019 Volume 10, Issue 6

Abstract: Pseudomonas ... ...

Abstract	Pseudomonas aeruginosa
MeSH term(s)	Biofilms/growth & development ; Cystic Fibrosis/complications ; Epithelial Cells/microbiology ; Extracellular Vesicles/metabolism ; Gene Deletion ; Host-Pathogen Interactions ; Humans ; Klebsiella pneumoniae/genetics ; Klebsiella pneumoniae/metabolism ; Leucyl Aminopeptidase/metabolism ; Protease Inhibitors/pharmacology ; Pseudomonas Infections/metabolism ; Pseudomonas Infections/microbiology ; Pseudomonas aeruginosa/drug effects ; Pseudomonas aeruginosa/enzymology ; Pseudomonas aeruginosa/genetics
Chemical Substances	Protease Inhibitors ; Leucyl Aminopeptidase (EC 3.4.11.1)
Language	English
Publishing date	2019-11-19
Publishing country	United States
Document type	Journal Article
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.02548-19
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Differential Packaging Into Outer Membrane Vesicles Upon Oxidative Stress Reveals a General Mechanism for Cargo Selectivity.

Orench-Rivera, Nichole / Kuehn, Meta J

Frontiers in microbiology

2021 Volume 12, Page(s) 561863

Abstract: Selective cargo packaging into bacterial extracellular vesicles has been reported and implicated in many biological processes, however, the mechanism behind the selectivity has remained largely unexplored. In this study, proteomic analysis of outer ... ...

Abstract	Selective cargo packaging into bacterial extracellular vesicles has been reported and implicated in many biological processes, however, the mechanism behind the selectivity has remained largely unexplored. In this study, proteomic analysis of outer membrane (OM) and OM vesicle (OMV) fractions from enterotoxigenic
Language	English
Publishing date	2021-07-02
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2021.561863
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Article ; Online: The extracellular vesicle generation paradox: a bacterial point of view.

McMillan, Hannah M / Kuehn, Meta J

The EMBO journal

2021 Volume 40, Issue 21, Page(s) e108174

Abstract: All bacteria produce secreted vesicles that carry out a variety of important biological functions. These extracellular vesicles can improve adaptation and survival by relieving bacterial stress and eliminating toxic compounds, as well as by facilitating ... ...

Abstract	All bacteria produce secreted vesicles that carry out a variety of important biological functions. These extracellular vesicles can improve adaptation and survival by relieving bacterial stress and eliminating toxic compounds, as well as by facilitating membrane remodeling and ameliorating inhospitable environments. However, vesicle production comes with a price. It is energetically costly and, in the case of colonizing pathogens, it elicits host immune responses, which reduce bacterial viability. This raises an interesting paradox regarding why bacteria produce vesicles and begs the question as to whether the benefits of producing vesicles outweigh their costs. In this review, we discuss the various advantages and disadvantages associated with Gram-negative and Gram-positive bacterial vesicle production and offer perspective on the ultimate score. We also highlight questions needed to advance the field in determining the role for vesicles in bacterial survival, interkingdom communication, and virulence.
MeSH term(s)	Animals ; Extracellular Vesicles/chemistry ; Extracellular Vesicles/metabolism ; Gene Expression ; Gram-Negative Bacteria/genetics ; Gram-Negative Bacteria/growth & development ; Gram-Negative Bacteria/metabolism ; Gram-Negative Bacteria/pathogenicity ; Gram-Positive Bacteria/genetics ; Gram-Positive Bacteria/growth & development ; Gram-Positive Bacteria/metabolism ; Gram-Positive Bacteria/pathogenicity ; Host-Parasite Interactions/genetics ; Humans ; Immunity, Innate ; Microbial Viability/genetics ; Quorum Sensing/genetics ; Secretory Vesicles/chemistry ; Secretory Vesicles/metabolism ; Virulence ; Virulence Factors/genetics ; Virulence Factors/metabolism
Chemical Substances	Virulence Factors
Language	English
Publishing date	2021-10-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	586044-1
ISSN	1460-2075 ; 0261-4189
ISSN (online)	1460-2075
ISSN	0261-4189
DOI	10.15252/embj.2021108174
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Zs.A 1808: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Extracellular Vesicles and Bacteriophages: New Directions in Environmental Biocolloid Research.

Hicks, Ethan / Rogers, Nicholas M K / Hendren, Christine Ogilvie / Kuehn, Meta J / Wiesner, Mark R

Environmental science & technology

2023 Volume 57, Issue 44, Page(s) 16728–16742

Abstract: There is a long-standing appreciation among environmental engineers and scientists regarding the importance of biologically derived colloidal particles and their environmental fate. This interest has been recently renewed in considering bacteriophages ... ...

Abstract	There is a long-standing appreciation among environmental engineers and scientists regarding the importance of biologically derived colloidal particles and their environmental fate. This interest has been recently renewed in considering bacteriophages and extracellular vesicles, which are each poised to offer engineers unique insights into fundamental aspects of environmental microbiology and novel approaches for engineering applications, including advances in wastewater treatment and sustainable agricultural practices. Challenges persist due to our limited understanding of interactions between these nanoscale particles with unique surface properties and their local environments. This review considers these biological particles through the lens of colloid science with attention given to their environmental impact and surface properties. We discuss methods developed for the study of inert (nonbiological) particle-particle interactions and the potential to use these to advance our understanding of the environmental fate and transport of extracellular vesicles and bacteriophages.
MeSH term(s)	Bacteriophages ; Environment ; Extracellular Vesicles ; Colloids
Chemical Substances	Colloids
Language	English
Publishing date	2023-10-29
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	1520-5851
ISSN (online)	1520-5851
DOI	10.1021/acs.est.3c05041
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Article: Breaking the bilayer: OMV formation during environmental transitions.

Bonnington, Katherine E / Kuehn, Meta J

Microbial cell (Graz, Austria)

2017 Volume 4, Issue 2, Page(s) 64–66

Abstract: Gram-negative bacteria maintain the barrier properties of the outer membrane (OM) in a wide array of physiological conditions despite their inability to degrade lipopolysaccharide (LPS) and protein material present in the outer leaflet of the OM. Through ...

Abstract	Gram-negative bacteria maintain the barrier properties of the outer membrane (OM) in a wide array of physiological conditions despite their inability to degrade lipopolysaccharide (LPS) and protein material present in the outer leaflet of the OM. Through characterization of the native dynamics of outer membrane LPS change we recently described a mechanism in which these diderm organisms overcome this design flaw. In response to different environmental stimuli
Language	English
Publishing date	2017-02-03
Publishing country	Austria
Document type	Journal Article ; Comment
ZDB-ID	2814756-X
ISSN	2311-2638
ISSN	2311-2638
DOI	10.15698/mic2017.02.558
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Article ; Online: Secreted bacterial vesicles as good samaritans.

Kuehn, Meta J

Cell host & microbe

2012 Volume 12, Issue 4, Page(s) 392–393

Abstract: Bacteriodes fragilis and polysaccharide A capsular antigen (PSA) produced by this commensal bacteria can mediate immune tolerance in the gastrointestinal (GI) tract. When looking for naturally secreted forms of PSA, Shen et al. (2012) suprisingly found ... ...

Abstract	Bacteriodes fragilis and polysaccharide A capsular antigen (PSA) produced by this commensal bacteria can mediate immune tolerance in the gastrointestinal (GI) tract. When looking for naturally secreted forms of PSA, Shen et al. (2012) suprisingly found that PSA is packaged in outer membrane vesicles (OMVs), bacterial blebs with a disreputable past.
Language	English
Publishing date	2012-10-16
Publishing country	United States
Document type	Comment ; Journal Article
ZDB-ID	2278004-X
ISSN	1934-6069 ; 1931-3128
ISSN (online)	1934-6069
ISSN	1931-3128
DOI	10.1016/j.chom.2012.10.005
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Article ; Online: Outer Membrane Vesicle Production Facilitates LPS Remodeling and Outer Membrane Maintenance in Salmonella during Environmental Transitions.

Bonnington, Katherine E / Kuehn, Meta J

mBio

2016 Volume 7, Issue 5

Abstract: The ability of Gram-negative bacteria to carefully modulate outer membrane (OM) composition is essential to their survival. However, the asymmetric and heterogeneous structure of the Gram-negative OM poses unique challenges to the cell's successful ... ...

Abstract	The ability of Gram-negative bacteria to carefully modulate outer membrane (OM) composition is essential to their survival. However, the asymmetric and heterogeneous structure of the Gram-negative OM poses unique challenges to the cell's successful adaption to rapid environmental transitions. Although mechanisms to recycle and degrade OM phospholipid material exist, there is no known mechanism by which to remove unfavorable lipopolysaccharide (LPS) glycoforms, except slow dilution through cell growth. As all Gram-negative bacteria constitutively shed OM vesicles (OMVs), we propose that cells may utilize OMV formation as a way to selectively remove environmentally disadvantageous LPS species. We examined the native kinetics of OM composition during physiologically relevant environmental changes in Salmonella enterica, a well-characterized model system for activation of PhoP/Q and PmrA/B two-component systems (TCSs). In response to acidic pH, toxic metals, antimicrobial peptides, and lack of divalent cations, these TCSs modify the LPS lipid A and core, lengthen the O antigen, and upregulate specific OM proteins. An environmental change to PhoP/Q- and PmrA/B-activating conditions simultaneously induced the addition of modified species of LPS to the OM, downregulation of previously dominant species of LPS, greater OMV production, and increased OMV diameter. Comparison of the relative abundance of lipid A species present in the OM and the newly budded OMVs following two sets of rapid environmental shifts revealed the retention of lipid A species with modified phosphate moieties in the OM concomitant with the selective loss of palmitoylated species via vesiculation following exposure to moderately acidic environmental conditions. Importance: All Gram-negative bacteria alter the structural composition of LPS present in their OM in response to various environmental stimuli. We developed a system to track the native dynamics of lipid A change in Salmonella enterica serovar Typhimurium following an environmental shift to PhoP/Q- and PmrA/B-inducing conditions. We show that growth conditions influence OMV production, size, and lipid A content. We further demonstrate that the lipid A content of OMVs does not fit a stochastic model of content selection, revealing the significant retention of lipid A species containing covalent modifications that mask their 1- and 4'-phosphate moieties under host-like conditions. Furthermore, palmitoylation of the lipid A to form hepta-acylated species substantially increases the likelihood of its incorporation into OMVs. These results highlight a role for the OMV response in OM remodeling and maintenance processes in Gram-negative bacteria.
MeSH term(s)	Exosomes/metabolism ; Lipopolysaccharides/metabolism ; Salmonella typhimurium/physiology ; Stress, Physiological
Chemical Substances	Lipopolysaccharides
Language	English
Publishing date	2016-10-18
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.01532-16
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Article ; Online: Protective plant immune responses are elicited by bacterial outer membrane vesicles.

McMillan, Hannah M / Zebell, Sophia G / Ristaino, Jean B / Dong, Xinnian / Kuehn, Meta J

Cell reports

2021 Volume 34, Issue 3, Page(s) 108645

Abstract: Bacterial outer membrane vesicles (OMVs) perform a variety of functions in bacterial survival and virulence. In mammalian systems, OMVs activate immune responses and are exploited as vaccines. However, little work has focused on the interactions of OMVs ... ...

Abstract	Bacterial outer membrane vesicles (OMVs) perform a variety of functions in bacterial survival and virulence. In mammalian systems, OMVs activate immune responses and are exploited as vaccines. However, little work has focused on the interactions of OMVs with plant hosts. Here, we report that OMVs from Pseudomonas syringae and P. fluorescens activate plant immune responses that protect against bacterial and oomycete pathogens. OMV-mediated immunomodulatory activity from these species displayed different sensitivity to biochemical stressors, reflecting differences in OMV content. Importantly, OMV-mediated plant responses are distinct from those triggered by conserved bacterial epitopes or effector molecules alone. Our study shows that OMV-induced protective immune responses are independent of the T3SS and protein, but that OMV-mediated seedling growth inhibition largely depends on proteinaceous components. OMVs provide a unique opportunity to understand the interplay between virulence and host response strategies and add a new dimension to consider in host-microbe interactions.
MeSH term(s)	Bacterial Outer Membrane Proteins/immunology ; Immunity/immunology ; Plant Immunity/immunology
Chemical Substances	Bacterial Outer Membrane Proteins
Language	English
Publishing date	2021-01-20
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2020.108645
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