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Article ; Online: RNA Extraction from Gram-Positive Bacteria Membrane Vesicles Using a Polymer-Based Precipitation Method.

Briaud, Paul / Carroll, Ronan K

Methods in molecular biology (Clifton, N.J.)

2024 Volume 2741, Page(s) 3–10

Abstract: Investigations into the biological role and composition of bacterial extracellular vesicles have grown in popularity in recent years. Vesicles perform a variety of functions during interactions with eukaryotic host cells, ranging from antibiotic ... ...

Abstract	Investigations into the biological role and composition of bacterial extracellular vesicles have grown in popularity in recent years. Vesicles perform a variety of functions during interactions with eukaryotic host cells, ranging from antibiotic resistance to immune modulation. It is necessary to isolate vesicles in order to understand their biological functions. Here we describe a polymer-based precipitation method allowing high-yield isolation of extracellular vesicles and their cargo RNA from the Gram-positive bacterium Staphylococcus aureus.
MeSH term(s)	Humans ; Membranes ; Gram-Positive Bacteria ; Extracellular Vesicles ; Staphylococcal Infections ; RNA
Chemical Substances	RNA (63231-63-0)
Language	English
Publishing date	2024-01-13
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3565-0_1
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Article ; Online: Extracellular Vesicle Biogenesis and Functions in Gram-Positive Bacteria.

Briaud, Paul / Carroll, Ronan K

Infection and immunity

2020 Volume 88, Issue 12

Abstract: Extracellular vesicles (EVs) are membrane-derived lipid bilayers secreted by bacteria and eukaryotic cells. Bacterial membrane vesicles were discovered over 60 years ago and have been extensively studied in Gram-negative bacteria. During their production, ...

Abstract	Extracellular vesicles (EVs) are membrane-derived lipid bilayers secreted by bacteria and eukaryotic cells. Bacterial membrane vesicles were discovered over 60 years ago and have been extensively studied in Gram-negative bacteria. During their production, EVs are loaded with proteins, nucleic acids, and various compounds that are subsequently released into the environment. Depending on the packaged cargo, EVs have a broad spectrum of action and are involved in pathogenesis, antibiotic resistance, nutrient uptake, and nucleic acid transfer. Due to differences in cell wall structure, EVs in Gram-positive bacteria have been disregarded for decades, and our understanding of their biogenesis and host cell interaction is incomplete. Recently, studies on bacteria such as
MeSH term(s)	Animals ; Bacillus subtilis/metabolism ; Bioengineering ; Biological Transport/genetics ; Extracellular Vesicles/chemistry ; Extracellular Vesicles/immunology ; Extracellular Vesicles/metabolism ; Gram-Positive Bacteria/genetics ; Gram-Positive Bacteria/metabolism ; Humans ; Inflammation/immunology ; Inflammation/metabolism ; Inflammation/microbiology ; Mycobacterium/metabolism ; Staphylococcal Infections/immunology ; Staphylococcal Infections/metabolism ; Staphylococcus aureus/metabolism ; Streptococcus/metabolism ; Vaccines
Chemical Substances	Vaccines
Language	English
Publishing date	2020-11-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.00433-20
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Article: How Bacterial Adaptation to Cystic Fibrosis Environment Shapes Interactions Between

Camus, Laura / Briaud, Paul / Vandenesch, François / Moreau, Karen

Frontiers in microbiology

2021 Volume 12, Page(s) 617784

Abstract: Pseudomonas ... ...

Abstract	Pseudomonas aeruginosa
Language	English
Publishing date	2021-03-03
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2021.617784
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Article ; Online: The Small Protein ScrA Influences Staphylococcus aureus Virulence-Related Processes via the SaeRS System.

Wittekind, Marcus A / Briaud, Paul / Smith, Jayanna L / Tennant, Julia R / Carroll, Ronan K

Microbiology spectrum

2023 Volume 11, Issue 3, Page(s) e0525522

Abstract: Staphylococcus aureus is a Gram-positive commensal and opportunistic pathogen able to cause diseases ranging from mild skin infections to life-threatening endocarditis and toxic shock syndrome. The ability to cause such an array of diseases is due to the ...

Abstract	Staphylococcus aureus is a Gram-positive commensal and opportunistic pathogen able to cause diseases ranging from mild skin infections to life-threatening endocarditis and toxic shock syndrome. The ability to cause such an array of diseases is due to the complex S. aureus regulatory network controlling an assortment of virulence factors, including adhesins, hemolysins, proteases, and lipases. This regulatory network is controlled by both protein and RNA elements. We previously identified a novel regulatory protein called ScrA, which, when overexpressed, leads to the increased activity and expression of the SaeRS regulon. In this study, we further explore the role of ScrA and examine the consequences to the bacterial cell of
MeSH term(s)	Animals ; Mice ; Virulence ; Staphylococcus aureus/metabolism ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Transcription Factors/metabolism ; Virulence Factors/metabolism ; Staphylococcal Infections/microbiology
Chemical Substances	Bacterial Proteins ; Transcription Factors ; Virulence Factors
Language	English
Publishing date	2023-05-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2807133-5
ISSN	2165-0497 ; 2165-0497
ISSN (online)	2165-0497
ISSN	2165-0497
DOI	10.1128/spectrum.05255-22
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Article: Mixed Populations and Co-Infection: Pseudomonas aeruginosa and Staphylococcus aureus.

Camus, Laura / Briaud, Paul / Vandenesch, François / Doléans-Jordheim, Anne / Moreau, Karen

Advances in experimental medicine and biology

2022 Volume 1386, Page(s) 397–424

Abstract: The human pathogens Pseudomonas aeruginosa and Staphylococcus aureus are frequently co-isolated from chronic wounds or cystic fibrosis patient airways. Clinical studies analysing the impact of co-infection on patient clinical outcomes lead to ... ...

Abstract	The human pathogens Pseudomonas aeruginosa and Staphylococcus aureus are frequently co-isolated from chronic wounds or cystic fibrosis patient airways. Clinical studies analysing the impact of co-infection on patient clinical outcomes lead to contradictory results. However, laboratory approaches suggest that the two pathogens co-colonize the same infection niches and form a mixed-species biofilm, therefore favouring their resistance to antibiotics and immune response. In parallel, many recent studies have focused on the different interactions between the two bacterial species. It has long been recognized that P. aeruginosa usually outcompetes S. aureus, and the molecular mechanisms involved in this state of bacterial competition are now well understood. However, several recent studies show that interactions between P. aeruginosa and S. aureus can be diverse and evolve over time. Thus, many CF isolates of P. aeruginosa and S. aureus can coexist and develop cooperative behaviours. In this chapter, we will provide an overview of the current knowledge on the mixed populations of P. aeruginosa and S. aureus, from their mechanisms of establishment to their impacts on bacterial physiology and clinical outcomes.
MeSH term(s)	Humans ; Staphylococcus aureus/physiology ; Pseudomonas aeruginosa/physiology ; Coinfection/microbiology ; Staphylococcal Infections/microbiology ; Biofilms ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use
Chemical Substances	Anti-Bacterial Agents
Language	English
Publishing date	2022-10-18
Publishing country	United States
Document type	Journal Article
ISSN	2214-8019 ; 0065-2598
ISSN (online)	2214-8019
ISSN	0065-2598
DOI	10.1007/978-3-031-08491-1_15
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Article ; Online: The novel protein ScrA acts through the SaeRS two-component system to regulate virulence gene expression in Staphylococcus aureus.

Wittekind, Marcus A / Frey, Andrew / Bonsall, Abigail E / Briaud, Paul / Keogh, Rebecca A / Wiemels, Richard E / Shaw, Lindsey N / Carroll, Ronan K

Molecular microbiology

2022 Volume 117, Issue 5, Page(s) 1196–1212

Abstract: Staphylococcus aureus is a Gram-positive commensal that can also cause a variety of infections in humans. S. aureus virulence factor gene expression is under tight control by a complex regulatory network, which includes, sigma factors, sRNAs, and two- ... ...

Abstract	Staphylococcus aureus is a Gram-positive commensal that can also cause a variety of infections in humans. S. aureus virulence factor gene expression is under tight control by a complex regulatory network, which includes, sigma factors, sRNAs, and two-component systems (TCS). Previous work in our laboratory demonstrated that overexpression of the sRNA tsr37 leads to an increase in bacterial aggregation. Here, we demonstrate that the clumping phenotype is dependent on a previously unannotated 88 amino acid protein encoded within the tsr37 sRNA transcript (which we named ScrA for S. aureus clumping regulator A). To investigate the mechanism of action of ScrA we performed proteomics and transcriptomics in a ScrA overexpressing strain and show that a number of surface adhesins are upregulated, while secreted proteases are downregulated. Results also showed upregulation of the SaeRS TCS, suggesting that ScrA is influencing SaeRS activity. Overexpression of ScrA in a saeR mutant abrogates the clumping phenotype confirming that ScrA functions via the Sae system. Finally, we identified the ArlRS TCS as a positive regulator of scrA expression. Collectively, our results show that ScrA is an activator of the SaeRS system and suggests that ScrA may act as an intermediary between the ArlRS and SaeRS systems.
MeSH term(s)	Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Gene Expression ; Gene Expression Regulation, Bacterial/genetics ; Humans ; Protein Kinases/metabolism ; RNA, Small Untranslated/metabolism ; Staphylococcal Infections ; Staphylococcus aureus/metabolism ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Virulence/genetics
Chemical Substances	Bacterial Proteins ; RNA, Small Untranslated ; Transcription Factors ; Protein Kinases (EC 2.7.-)
Language	English
Publishing date	2022-04-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	619315-8
ISSN	1365-2958 ; 0950-382X
ISSN (online)	1365-2958
ISSN	0950-382X
DOI	10.1111/mmi.14901
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Article ; Online: Global Annotation, Expression Analysis, and Stability of Candidate sRNAs in Group B Streptococcus.

Keogh, Rebecca A / Spencer, Brady L / Sorensen, Hailee M / Zapf, Rachel L / Briaud, Paul / Bonsall, Abigail E / Doran, Kelly S / Carroll, Ronan K

mBio

2021 Volume 12, Issue 6, Page(s) e0280321

Abstract: Small, noncoding RNAs (sRNAs) are being increasingly identified as important regulatory molecules in prokaryotes. Due to the prevalence of next-generation sequencing-based techniques, such as RNA sequencing (RNA-seq), there is potential for increased ... ...

Abstract	Small, noncoding RNAs (sRNAs) are being increasingly identified as important regulatory molecules in prokaryotes. Due to the prevalence of next-generation sequencing-based techniques, such as RNA sequencing (RNA-seq), there is potential for increased discovery of sRNAs within bacterial genomes; however, these elements are rarely included in annotation files. Consequently, expression values for sRNAs are omitted from most transcriptomic analyses, and mechanistic studies have lagged behind those of protein regulators in numerous bacteria. Two previous studies have identified sRNAs in the human pathogen group B Streptococcus (GBS). Here, we utilize the data from these studies to create updated genome annotation files for the model GBS strains NEM316 and COH1. Using the updated COH1 annotation file, we reanalyze publicly available GBS RNA-seq whole-transcriptome data from GenBank to monitor GBS sRNA expression under a variety of conditions and genetic backgrounds. This analysis generated expression values for 232 putative sRNAs that were overlooked in previous transcriptomic analyses in 21 unique comparisons. To demonstrate the utility of these data, we identify an sRNA that is upregulated during vaginal colonization and demonstrate that overexpression of this sRNA leads to increased bacterial invasion into host epithelial cells. Finally, to monitor RNA degradation, we perform a transcript stability assay to identify highly stable sRNAs and compare stability profiles of sRNA- and protein-coding genes. Collectively, these data provide a wealth of transcriptomic data for putative sRNAs in GBS and a platform for future mechanistic studies.
MeSH term(s)	Gene Expression Profiling ; Gene Expression Regulation, Bacterial ; Genome, Bacterial ; Humans ; RNA Stability ; RNA, Bacterial/chemistry ; RNA, Bacterial/genetics ; RNA, Bacterial/metabolism ; RNA, Small Untranslated/chemistry ; RNA, Small Untranslated/genetics ; RNA, Small Untranslated/metabolism ; Streptococcal Infections/microbiology ; Streptococcus agalactiae/chemistry ; Streptococcus agalactiae/genetics ; Streptococcus agalactiae/metabolism
Chemical Substances	RNA, Bacterial ; RNA, Small Untranslated
Language	English
Publishing date	2021-11-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.02803-21
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Article ; Online: Coexistence with Pseudomonas aeruginosa alters Staphylococcus aureus transcriptome, antibiotic resistance and internalization into epithelial cells.

Briaud, Paul / Camus, Laura / Bastien, Sylvère / Doléans-Jordheim, Anne / Vandenesch, François / Moreau, Karen

Scientific reports

2019 Volume 9, Issue 1, Page(s) 16564

Abstract: Cystic fibrosis (CF) is the most common life-threatening genetic disease among Caucasians. CF patients suffer from chronic lung infections due to the presence of thick mucus, caused by cftr gene dysfunction. The two most commonly found bacteria in the ... ...

Abstract	Cystic fibrosis (CF) is the most common life-threatening genetic disease among Caucasians. CF patients suffer from chronic lung infections due to the presence of thick mucus, caused by cftr gene dysfunction. The two most commonly found bacteria in the mucus of CF patients are Staphylococcus aureus and Pseudomonas aeruginosa. It is well known that early-infecting P. aeruginosa strains produce anti-staphylococcal compounds and inhibit S. aureus growth. More recently, it has been shown that late-infecting P. aeruginosa strains develop commensal-like/coexistence interaction with S. aureus. The aim of this study was to decipher the impact of P. aeruginosa strains on S. aureus. RNA sequencing analysis showed 77 genes were specifically dysregulated in the context of competition and 140 genes in the context of coexistence in the presence of P. aeruginosa. In coexistence, genes encoding virulence factors and proteins involved in carbohydrates, lipids, nucleotides and amino acids metabolism were downregulated. On the contrary, several transporter family encoding genes were upregulated. In particular, several antibiotic pumps belonging to the Nor family were upregulated: tet38, norA and norC, leading to an increase in antibiotic resistance of S. aureus when exposed to tetracycline and ciprofloxacin and an enhanced internalization rate within epithelial pulmonary cells. This study shows that coexistence with P. aeruginosa affects the S. aureus transcriptome and virulence.
MeSH term(s)	A549 Cells ; Cystic Fibrosis/microbiology ; Drug Resistance, Microbial/genetics ; Endocytosis ; Epithelial Cells/microbiology ; Gene Expression Regulation, Bacterial ; Genes, Bacterial ; Humans ; Pseudomonas aeruginosa/isolation & purification ; Pseudomonas aeruginosa/physiology ; Reproducibility of Results ; Staphylococcus aureus/genetics ; Staphylococcus aureus/isolation & purification ; Transcriptome/genetics
Language	English
Publishing date	2019-11-12
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-52975-z
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Article ; Online: The Small RNA Teg41 Is a Pleiotropic Regulator of Virulence in Staphylococcus aureus.

Briaud, Paul / Zapf, Rachel L / Mayher, Ariana D / McReynolds, Aubrey K G / Frey, Andrew / Sudnick, Emily G / Wiemels, Richard E / Keogh, Rebecca A / Shaw, Lindsey N / Bose, Jeffrey L / Carroll, Ronan K

Infection and immunity

2022 Volume 90, Issue 11, Page(s) e0023622

Abstract: Previously, our group demonstrated a role for the small RNA (sRNA) Teg41 in regulating production of the alpha phenol-soluble modulin toxins (αPSMs) in Staphylococcus aureus. Overexpressing Teg41 increased αPSM production while deleting the 3' end of ... ...

Abstract	Previously, our group demonstrated a role for the small RNA (sRNA) Teg41 in regulating production of the alpha phenol-soluble modulin toxins (αPSMs) in Staphylococcus aureus. Overexpressing Teg41 increased αPSM production while deleting the 3' end of Teg41 (Teg41Δ3' strain) resulted in a decrease in αPSM production, reduced hemolytic activity of S. aureus culture supernatants, and attenuated virulence in a murine abscess model of infection. In this study, we further explore the attenuation of virulence in the Teg41Δ3' strain. Using both localized and systemic models of infection, we demonstrate that the Teg41Δ3' strain is more severely attenuated than an ΔαPSM mutant, strongly suggesting that Teg41 influences more than the αPSMs. Proteomic and transcriptomic analysis of the wild-type and Teg41Δ3' strains reveals widespread alterations in transcript abundance and protein production in the absence of Teg41, confirming that Teg41 has pleiotropic effects in the cell. We go on to investigate the molecular mechanism underlying Teg41-mediated gene regulation. Surprisingly, results demonstrate that certain Teg41 target genes, including the αPSMs and βPSMs, are transcriptionally altered in the Teg41Δ3' strain, while other targets, specifically
MeSH term(s)	Mice ; Animals ; Staphylococcus aureus ; Virulence ; RNA/metabolism ; Proteomics ; Gene Expression Regulation, Bacterial ; Virulence Factors/genetics ; Virulence Factors/metabolism ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Staphylococcal Infections/metabolism
Chemical Substances	RNA (63231-63-0) ; Virulence Factors ; Bacterial Proteins
Language	English
Publishing date	2022-10-10
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/iai.00236-22
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Article ; Online: Trophic cooperation promotes bacterial survival of Staphylococcus aureus and Pseudomonas aeruginosa.

Camus, Laura / Briaud, Paul / Bastien, Sylvère / Elsen, Sylvie / Doléans-Jordheim, Anne / Vandenesch, François / Moreau, Karen

The ISME journal

2020 Volume 14, Issue 12, Page(s) 3093–3105

Abstract: In the context of infection, Pseudomonas aeruginosa and Staphylococcus aureus are frequently co-isolated, particularly in cystic fibrosis (CF) patients. Within lungs, the two pathogens exhibit a range of competitive and coexisting interactions. In the ... ...

Abstract	In the context of infection, Pseudomonas aeruginosa and Staphylococcus aureus are frequently co-isolated, particularly in cystic fibrosis (CF) patients. Within lungs, the two pathogens exhibit a range of competitive and coexisting interactions. In the present study, we explored the impact of S. aureus on the physiology of P. aeruginosa in the context of coexistence. Transcriptomic analyses showed that S. aureus significantly and specifically affects the expression of numerous genes involved in P. aeruginosa carbon and amino acid metabolism. In particular, 65% of the strains presented considerable overexpression of the genes involved in the acetoin catabolic (aco) pathway. We demonstrated that acetoin is (i) produced by clinical S. aureus strains, (ii) detected in sputa from CF patients and (iii) involved in P. aeruginosa's aco system induction. Furthermore, acetoin is catabolized by P. aeruginosa, a metabolic process that improves the survival of both pathogens by providing a new carbon source for P. aeruginosa and avoiding the toxic accumulation of acetoin on S. aureus. Due to its beneficial effects on both bacteria, acetoin catabolism could testify to the establishment of trophic cooperation between S. aureus and P. aeruginosa in the CF lung environment, thus promoting their persistence.
MeSH term(s)	Biofilms ; Humans ; Microbial Interactions ; Pseudomonas Infections ; Pseudomonas aeruginosa/genetics ; Staphylococcal Infections ; Staphylococcus aureus/genetics
Language	English
Publishing date	2020-08-19
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2406536-5
ISSN	1751-7370 ; 1751-7362
ISSN (online)	1751-7370
ISSN	1751-7362
DOI	10.1038/s41396-020-00741-9
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