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Staphylococcus aureus surface LC-MS/MS analysis

ABSTRACT: Staphylococcus aureus causes treatment-resistant infections, to which biofilm formation and bacterial persistence contribute greatly. This study aims at the large-scale characterization of mechanisms behind its prolonged persistence in hostile environments and capability to evade host defenses. We carried out the analysis of surface-exposed proteins in S. aureus biofilms prepared from two types of inocula; metabolically active, levofloxacin-susceptible culture and stationary phase, levofloxacin-tolerant culture. Age-synchronized planktonic control cultures provided a reference to facilitate the identification of growth mode-specific surface-expression patterns.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Staphylococcus Aureus

SUBMITTER: Tuula Nyman

LAB HEAD: Tuula Nyman

PROVIDER: PXD016344 | Pride | 2020-01-20

REPOSITORIES: Pride

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Publications

Growth Mode and Physiological State of Cells Prior to Biofilm Formation Affect Immune Evasion and Persistence of <i>Staphylococcus aureus</i>.

Savijoki Kirsi K Miettinen Ilkka I Nyman Tuula A TA Kortesoja Maarit M Hanski Leena L Varmanen Pekka P Fallarero Adyary A

Microorganisms 20200112 1

The present study investigated <i>Staphylococcus aureus</i> ATCC25923 surfaceomes (cell surface proteins) during prolonged growth by subjecting planktonic and biofilm cultures (initiated from exponential or stationary cells) to label-free quantitative surfaceomics and phenotypic confirmations. The abundance of adhesion, autolytic, hemolytic, and lipolytic proteins decreased over time in both growth modes, while an opposite trend was detected for many tricarboxylic acid (TCA) cycle, reactive oxyg ...[more]

PMID: 31940921

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Surfaceome and exoproteome analysis of Staphylococcus aureus and Pseudomonas aeruginosa mono- and co-culture biofilms

Project description:Staphylococcus aureus and Pseudomonas aeruginosa are bacterial pathogens that have been shown to co-exist in biofilms related to numerous infections. Although the interaction between these two species is competitive, both partially benefit from the coexistence. In this study, we exhaustively characterized the interaction between Staphylococcus aureus and Pseudomonas aeruginosa by utilizing a proteomics approach, individually targeting the surface-associated proteins (surfaceome), and proteins secreted or otherwise liberated to the extracellular space (exoproteome). To that end, the conditions to co-culture S. aureus and P. aeruginosa in vitro were optimized and a high-resolution proteomics approach was applied to compare surface-associated and extracellular protein profiles between mono- and co-cultured biofilms.

2021-09-09 | PXD023445 | Pride

Phenotype and expression profile analysis of Staphylococcus aureus biofilms and planktonic cells in response to Licochalcone A

Project description:Staphylococcus aureus (S. aureus) is one of the most important pathogens in humans and animals. The formation of S. aureus biofilm is considered an important mechanism of antimicrobial resistance. Therefore, finding effective drugs against the biofilm produced by S. aureus has been a high priority. Licochalcone A, a natural plant product, was reported to have antibacterial activities and showed good activity against all 21 tested strains of S. aureus biofilm and planktonic cells. To detect the possible molecular mechanism of Licochalcone A against S. aureus biofillm or planktonic cells, Affymetrix GeneChips were used to determine the global comparative transcription of S. aureus biofilm and planktonic cells triggered by treatment with sub-bactericidal and sub-inhibitory concentrations of Licochalcone A, respectively. Staphylococcus aureus planktonic cells and biofilm were exposed for 60 minutes to Licochalcone A at concentration of 2 M-NM-<g/ml (1/2M-CM-^W MIC) and 64 M-NM-<g/ml (4M-CM-^W MIBC), respectively. 4 samples including 4 control samples are analyzed.

2014-07-01 | E-GEOD-58938 | biostudies-arrayexpress

Phenotype and RNA-Seq-Based Transcriptome Profiling of Staphylococcus aureus biofilms in response to Tea tree oil

Project description:RNA sequencing (RNA-Seq) was used in our study to elucidate the mechanism of Tea tree oil (TTO) as a potential antibacterial agent to evaluate differentially expressed genes and functional network analysis in S. aureus ATCC 29213 biofilms. Staphylococcus aureus biofilm cells were exposed for 60 minutes to TTO at concentration of 1/2ÃMBIC (1 mg/ml).2 samples including 2 control samples are analyzed.

2016-08-19 | E-GEOD-85787 | biostudies-arrayexpress

The type VII secretion system protects Staphylococcus aureus against host-specific fatty acids

Project description:The Staphylococcus aureus type VII secretion system (T7SS) exports several proteins that are pivotal for bacterial virulence. The mechanisms underlying T7SS-mediated staphylococcal survival during infection are nevertheless unclear. Here we show that EsxC, a small secreted effector implicated in bacterial persistence, contributes to S. aureus membrane architecture and fluidity. Interestingly, isogenic mutants lacking EsxC, T7SS effectors (EsxA or EsxB), and the membrane-bound EssC are more sensitive to killing by the host-specific fatty acid, linoleic acid (LA), compared to the wild-type. We demonstrate that LA induces more cell membrane damage in the T7SS mutants, although they do not bind differentially to LA. Membrane lipid profiles show that T7SS mutants are also less able to incorporate LA into their membrane phospholipids. Proteomics analyses of wild-type and mutant cell fractions reveal that, in addition to compromising membranes, T7SS defects readily induce bacterial stress and hamper their response to LA challenge. Together, our findings indicate that T7SS is crucial for S. aureus membrane integrity and homeostasis, which is critical when bacteria encounter antimicrobial fatty acids.

2021-09-08 | PXD013081 | Pride

The Staphylococcus aureus Host Cell Genetic Adhesion Network

Project description:Adhesion of Staphylococcus aureus to the host’s skin and mucosae enables asymptomatic colonization and the establishment of infections. This process is facilitated by cell wall-anchored (CWA) adhesins that are covalently attached to the bacterial cell wall and exhibit affinity for host ligands. To gain insight into the cellular factors contributing to adhesion, we developed a sensitive and robust assay to enable the large-scale profiling of S. aureus adhesion to host ligands. Using this assay, we profiled a sequence defined transposon mutant library of the community-associated methicillin-resistant S. aureus (CA-MRSA) isolate USA300, to identify mutant strains with attenuated adhesion to human derived keratin, fibronectin and fibrinogen. Overall, we identified 21 adhesion defective mutants and we used this information to construct ‘The S. aureus Genetic Adhesion Network’. Importantly, we identified a core gene set involved in adhesion to all three host ligands and we delineated unique genetic signatures. In addition, we demonstrate a central requirement for autolysin-mediated daughter cell separation in S. aureus CWA protein-mediated adhesion to host ligands, which contributes to virulence in murine skin and soft tissue infection. This comprehensive study will inform strategies to inhibit S. aureus host cell adhesion, potentially enabling the development of anti-adhesive therapeutics to complement traditional antibacterial chemotherapy.

2023-03-04 | PXD018514 | Pride

Secreted Factors from Staphylococcus aureus Biofilm and Planktonic Cultures Differentially Impact Human Keratinocytes in vitro

Project description:Interactions between human keratinocytes and secreted factors from Staphylococcus aureus biofilm were investigated using microarray analysis. Relative to control cells, biofilm-secreted factors upregulated cytokine and chemokine genes in keratinocytes. Genes associated with DNA damage and oxidative stress were also induced in keratinocytes treated with secreted factors from S. aureus. Here we show that secreted factors from S. aureus biofilm cultures differentially impact several aspects of wound healing processes. Human keratinocytes were grown in co-culture with mature S. aureus biofilms for 24 hours. Keratinocytes exposed to S. aureus biofilm were analyzed in quadruplicate. Control cells were also analyzed in quadruplicate. Dye-swaps were performed.

2011-08-30 | E-GEOD-24370 | biostudies-arrayexpress

Proteome of Staphylococcus aureus biofilm changes signifi-cantly with aging

Project description:Staphylococcus aureus is a notorious biofilm-producing pathogen that is frequently isolated from implantable medical device infections. As biofilm ages, it becomes more tolerant to antimicrobial treatment leading to treatment failure and necessitating the costly removal of infected devices. In this study, we investigated what changes occur in the proteome of S. aureus biofilm grown for 3-days and 12-days in comparison with 24 h planktonic showed that proteins associated with bi-osynthetic processes, ABC transporter pathway, virulence proteins, and shikimate kinase pathway were significantly upregulated in 3-day biofilm, while proteins associated with sugar transporter, degradation, and stress response were downregulated. In 12-day biofilms, proteins associated with peptidoglycan biosynthesis, sugar transporters, and stress responses were upregulated, whereas proteins associated with ABC transporters, DNA replication, and adhesion proteins were down-regulated. Furthermore, we observed significant variations in the formation of biofilms result from changes in the level of metabolic activity in the different growth mode of biofilms that could be a significant factor of S. aureus biofilm maturation and persistence. Collectively, potential marker proteins were identified and further characterized to understand their exact role in S. aureus biofilm development which may shed light on possible new therapeutic regimes in the treatment of biofilm-related implant-associated infections.

2022-08-12 | PXD033499 | Pride

Co-culture of C. albicans with bacteria

Project description:Transcriptional profiling of C. albicans when monocultured in suspension culture or in biofilms compared to when co-cultured with C. perfringens in suspension culture, or with E. coli, K. pneumoniae, E. faecalis, C. perfringens, or B. fragilis in biofilms, or in the presence of N-acetylglucosamine in biofilms. At least 2 replicates for each condition except for biofilms +N-acetylglucosamine, some experiments are two-condition experiments: C. albicans +/- bacteria vs. C. albicans. Some experiments multiple condition: C. albicans +/- bacteria vs. mixed reference of all conditions in that experiment.

2014-03-03 | E-GEOD-55026 | biostudies-arrayexpress

GdpS contributes to Staphylococcus aureus biofilm formation by regulation of eDNA release

Project description:In Staphylococcus aureus, the role of the GGDEF domain containing protein GdpS remains poorly understood. Previous studies reported that gdpS mutant strains had decreased biofilm formation due to changes in icaADBC expression that were independent of cyclic-di-GMP levels. We deleted gdpS in three unrelated S. aureus isolates, and analyzed the resultant mutants for alterations in biofilm formation, metabolism and transcription. Dynamic imaging during biofilm development showed that GdpS inhibited early biofilm formation in only two out of the three strains examined, without affecting bacterial survival. However, quantification of biofilm formation using crystal violet staining revealed that inactivation of gdpS affected biofilm formation in all three studied strains. Extraction of metabolites from S. aureus cells confirmed the absence of cyclic-di-GMP, suggesting that biofilm formation in this species differs from that in other Gram-positive organisms. In addition, targeted mutagenesis demonstrated that the GGDEF domain was not required for GdpS activity. Transcriptomic analysis revealed that the vast majority of GGDEF-regulated genes were involved in virulence, metabolism, cell wall biogenesis and eDNA release. Finally, expression of lrgAB or deletion of cidABC in a strain lacking gdpS confirmed the role of GdpS on regulation of eDNA production that occurred without an increase in cell autolysis. In summary, S. aureus GdpS contributes to cell-to-cell interactions during early biofilm formation by influencing expression of lrgAB and cidABC mediated eDNA release. We conclude that GdpS acts as a negative regulator of eDNA release. Three strains UAMS-1, SA113 and SA564 were used in this study to compare wt with gdpS mutant after 5 hours of growth in static conditions (biofilm formation).

2014-06-30 | E-GEOD-48656 | biostudies-arrayexpress

Global transcriptome analysis of Staphylococcus aureus biofilms in response to innate immune cells

Project description:S. aureus biofilms are associated with the organism's ability to cause disease. Biofilm associated bacteria must cope with the host's innate immune system. We used commercially available Affymetrix S. aureus GeneChips to compare the gene expression properties of 4 and 6 day established biofilms following short (1 hr)- and long (24 hr)- term exposure to macrophages and neutrophils. S. aureus strain USA300 LAC biofilms where formed for 4 or 6 days. Established biofilms were then exposed to macrophages for 1 or 24 hr. Alternatively, biofilms were exposed to neutrophils for 1 or 4 hr. Total bacterial RNA was isolated and subjected to GeneChip hybridization and analysis. We sought to determine the regulatory effects of Macrophages and Neutrophils on established S. aureus biofilms.

2013-12-01 | E-GEOD-50675 | biostudies-arrayexpress

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