Protein homeostasis impairment affects phenotypic heterogeneity of biofilms
Ontology highlight
ABSTRACT:
ORGANISM(S): Bacillus subtilis
PROVIDER: GSE276606 | GEO | 2025/07/16
REPOSITORIES: GEO
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Project description:Biofilms are matrix-encased microbial communities that increase the environmental fitness and infectivity of many human pathogens including Vibrio cholerae. Biofilm matrix assembly is essential for biofilm formation and function. Known components of the V. cholerae biofilm matrix are the polysaccharide VPS, matrix proteins RbmA, RbmC, Bap1, and extracellular DNA, but the majority of the protein composition is uncharacterized. This study comprehensively analyzed the biofilm matrix proteome and revealed the presence of outer membrane proteins (OMPs). Outer membrane vesicles (OMVs) were also present in the V. cholerae biofilm matrix and were associated with OMPs and many biofilm matrix proteins suggesting that they participate in biofilm matrix assembly. Consistent with this, OMVs had the capability to alter biofilm structural properties depending on their composition. OmpU was the most prevalent OMP in the matrix, and its absence altered biofilm architecture by increasing VPS production. Using single-cell force spectroscopy, we further showed that OmpU, the matrix proteins RbmA, RbmC, and Bap1, and VPS contribute to cell-surface adhesion forces, which are critical for biofilm formation. Our findings provide new insights into the molecular mechanisms underlying biofilm matrix assembly in V. cholerae, which may open up new opportunities to develop inhibitors that specifically alter biofilm matrix properties and, thus, affect either the environmental survival or pathogenesis of Vibrio cholerae.
2023-07-14 | MSV000092446 | MassIVE
Project description:Biofilms are matrix-encased microbial communities that increase the environmental fitness and infectivity of many human pathogens including Vibrio cholerae. Biofilm matrix assembly is essential for biofilm formation and function. Known components of the V. cholerae biofilm matrix are the polysaccharide VPS, matrix proteins RbmA, RbmC, Bap1, and extracellular DNA, but the majority of the protein composition is uncharacterized. This study comprehensively analyzed the biofilm matrix proteome and revealed the presence of outer membrane proteins (OMPs). Outer membrane vesicles (OMVs) were also present in the V. cholerae biofilm matrix and were associated with OMPs and many biofilm matrix proteins suggesting that they participate in biofilm matrix assembly. Consistent with this, OMVs had the capability to alter biofilm structural properties depending on their composition. OmpU was the most prevalent OMP in the matrix, and its absence altered biofilm architecture by increasing VPS production. Single-cell force spectroscopy revealed that proteins critical for biofilm formation, OmpU, the matrix proteins RbmA, RbmC, Bap1, and VPS, contribute to cell-surface adhesion forces at differing efficiency, with VPS showing the highest efficiency whereas Bap1 showing the lowest efficiency. Our findings provide new insights into the molecular mechanisms underlying biofilm matrix assembly in V. cholerae, which may provide new opportunities to develop inhibitors that specifically alter biofilm matrix properties and, thus, affect either the environmental survival or pathogenesis of V. cholerae.
2023-12-12 | MSV000093650 | MassIVE
Project description:Biofilms are surface-adhered bacterial communities encased in an extracellular matrix composed of polysaccharides, proteins, and extracelluar (e)DNA, with eDNA being required for the formation and integrity of biofilms. Here we demonstrate that the spatial and temporal release of eDNA is regulated by BfmR, a regulator essential for Pseudomonas aeruginosa biofilm development. The expression of bfmR coincided with localized cell death and DNA release, with high eDNA concentrations localized to the outer part of microcolonies in the form of a ring and as a cap on small clusters. Additionally, eDNA release and cell lysis increased significantly following bfmR inactivation. Genome-wide transcriptional profiling indicated that bfmR was required for repression of genes associated with bacteriophage assembly and bacteriophage-mediated lysis. In order to determine which of these genes were directly regulated by BfmR, we utilized chromatin immunoprecipitation (ChIP) analysis to identify the promoter of PA0691, termed here phdA, encoding a previously undescribed homologue of the prevent-host-death (Phd) family of proteins. Lack of phdA expression coincided with impaired biofilm development, increased cell death and bacteriophage release, a phenotype comparable to ΔbfmR. Expression of phdA in ΔbfmR biofilms restored eDNA release, cell lysis, release of bacteriophages, and biofilm formation to wild type levels. Moreover, overexpression of phdA rendered P. aeruginosa resistant to lysis mediated by superinfective bacteriophage Pf4 which was only detected in biofilms. The expression of bfmR was stimulated by conditions resulting in membrane perturbation and cell lysis. Thus, we propose that BfmR regulates biofilm development by controlling bacteriophage-mediated lysis and thus, cell death and eDNA release, via PhdA. For biofilm growth experiments, three independent replicates of P. aeruginosa strains PAO1 and ΔbfmR were grown as biofilms in a flow-through system using a once-through continuous flow tube reactor system for biofilm sample collection and in flow cells (BioSurface Technologies) for the analysis of biofilm architecture as previously described (Sauer et al., 2002, Sauer et al., 2004, Petrova & Sauer, 2009). Cells were treated with RNAprotect (Qiagen), and total RNA was extracted using an RNeasy mini purification kit (Qiagen) per the manufacturer’s instructions. RNA quality and the presence of residual DNA were checked on an Agilent Bioanalyzer 2100 electrophoretic system pre- and post-DNase treatment. Ten micrograms of total RNA was used for cDNA synthesis, fragmentation, and labeling according to the Affymetrix GeneChip P. aeruginosa genome array expression analysis protocol. Sauer, K., A. K. Camper, G. D. Ehrlich, J. W. Costerton & D. G. Davies, (2002) Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J. Bacteriol. 184: 1140-1154. Sauer, K., M. C. Cullen, A. H. Rickard, L. A. H. Zeef, D. G. Davies & P. Gilbert, (2004) Characterization of nutrient-induced dispersion in Pseudomonas aeruginosa PAO1 biofilm. J. Bacteriol. 186: 7312-7326. Petrova, O. E. & K. Sauer, (2009) A novel signaling network essential for regulating Pseudomonas aeruginosa biofilm development. PLoS Pathogens 5: e1000668.
2011-06-01 | E-GEOD-28953 | biostudies-arrayexpress
Project description:Biofilms are surface-adhered bacterial communities encased in an extracellular matrix composed of polysaccharides, proteins, and extracelluar (e)DNA, with eDNA being required for the formation and integrity of biofilms. Here we demonstrate that the spatial and temporal release of eDNA is regulated by BfmR, a regulator essential for Pseudomonas aeruginosa biofilm development. The expression of bfmR coincided with localized cell death and DNA release, with high eDNA concentrations localized to the outer part of microcolonies in the form of a ring and as a cap on small clusters. Additionally, eDNA release and cell lysis increased significantly following bfmR inactivation. Genome-wide transcriptional profiling indicated that bfmR was required for repression of genes associated with bacteriophage assembly and bacteriophage-mediated lysis. In order to determine which of these genes were directly regulated by BfmR, we utilized chromatin immunoprecipitation (ChIP) analysis to identify the promoter of PA0691, termed here phdA, encoding a previously undescribed homologue of the prevent-host-death (Phd) family of proteins. Lack of phdA expression coincided with impaired biofilm development, increased cell death and bacteriophage release, a phenotype comparable to ΔbfmR. Expression of phdA in ΔbfmR biofilms restored eDNA release, cell lysis, release of bacteriophages, and biofilm formation to wild type levels. Moreover, overexpression of phdA rendered P. aeruginosa resistant to lysis mediated by superinfective bacteriophage Pf4 which was only detected in biofilms. The expression of bfmR was stimulated by conditions resulting in membrane perturbation and cell lysis. Thus, we propose that BfmR regulates biofilm development by controlling bacteriophage-mediated lysis and thus, cell death and eDNA release, via PhdA.
2011-06-01 | GSE28953 | GEO
Project description:The opportunistic pathogenic mold Aspergillus fumigatus is an increasing cause of morbidity and mortality in immunocompromised and, in part, immunocompetent patients. Like bacteria or yeast, A. fumigatus can grow in multicellular communities by the formation of a hyphal network encased in an extracellular matrix. Here, we describe the proteome and transcriptome of planktonic and biofilm-grown A. fumigatus mycelium after 24h and 48h. A biofilm- and time-dependent regulation of many proteins and genes of the primary metabolism indicates a developmental stage of the young biofilm at 24h, which demands energy. At a matured biofilm phase, metabolic activity seems to be reduced. However, genes encoding hydrophobins and proteins involved in the biosynthesis of secondary metabolites were significantly upregulated. In particular, proteins of the gliotoxin secondary metabolite gene cluster were induced in biofilm cultures. This was confirmed by RT-PCR and by detection of this immunologically active mycotoxin in culture supernatants using HPLC analysis. The enhanced production of gliotoxin by in vitro formed biofilms reported here may play also a significant role under in vivo conditions. It may confer A. fumigatus protection from the host immune system and also enable its survival and persistence in chronic lung infections such as aspergilloma. Comparison of biofilm and submers cultures at 24h and 48h after induction.
2010-07-01 | E-GEOD-19430 | biostudies-arrayexpress
Project description:Extracellular polymeric substances (EPS) produced by bacteria form a matrix for the complex three-dimensional architecture of biofilms. The EPS matrix of a biofilm is primarily composed of polysaccharides, proteins and extracellular DNA (eDNA). In addition to supporting the community structure, the EPS matrix protects bacterial biofilms from the environment. Specifically, the EPS matrix reduces the penetration of antimicrobial agents into the biofilm, thereby reducing their efficiency. Strategies for disrupting the formation of the EPS matrix can therefore lead to a more efficient use of antimicrobial agents. Here, we examine the biofilm-disrupting activity of vitamin C (sodium ascorbate). Quantitative proteomics analysis of vitamin C treated biofilms show that exposure to vitamin C inhibits quorum sensing and other regulatory mechanisms underpinning biofilm development. This results in inhibition of the EPS biosynthesis, and depletion of the polysaccharide component of the matrix. Once the EPS content is reduced beyond a critical point, bacterial cells get fully exposed to the medium, thereby experiencing a planktonic-like state. At this stage, the cells are more susceptible to killing, either by vitamin C-induced oxidative stress as reported here, or by other antibacterial compounds or treatments.
2017-12-19 | PXD007533 | Pride
Project description:To explain enhanced biofilm formation and increased dissemination of S. epidermidis in mixed-species biofilms, microarrays were used to explore differential gene expression of S. epidermidis in mixed-species biofilms. One sample from single species biofilm (S1) and mixed-species biofilm (SC2) were excluded from analyses for outliers. We observed upregulation (2.7%) and down regulation (6%) of S. epidermidis genes in mixed-species biofilms. Autolysis repressors lrgA and lrgB were down regulated 36-fold and 27-fold respectively and was associated with increased eDNA possibly due to enhanced autolysis in mixed-species biofilms. These data suggest that bacterial autolysis and release of eDNA in the biofilm matrix may be responsible for enhancement and dissemination of mixed-species biofilms of S. epidermidis and C. albicans.
2012-02-28 | GSE35438 | GEO
Project description:Tissue-penetrating nanobodies as drug-delivery vehicles have shown efficacy in treating hard-to-reach and -treat diseases. The present study reports nanobodies with application potential in the treatment of tuberculosis (TB), especially the antibiotic tolerant biofilms that form in TB granulomas. Using Mycobacterium marinum (Mmr) as a model pathogen, we identified the most abundant matrix proteins in cultured biofilms and granuloma biofilms from infected zebrafish, and used them as targets to produce synthetic nanobodies (sybodies). Surface-exposed proteins on Mmr biofilms were identified using a combination of in vitro cell surface biotinylation proteomics andex vivo proteomics on granulomas extracted from Mmr-infected adult zebrafish. We identified a total of 3080 and 41 mycobacterial biofilm matrix-associated proteins by in vitro and ex vivo proteomics, respectively. The molecular chaperones GroEL1 and GroEL2 were identified in both datasets, and fulfilled the criteria of ideal nanobody targets. We created sybodies against GroEL1 and GroEL2, and showed that they bind the intact in vitro and ex vivo Mmr-biofilms. Taken together, the present study reports a proof-of-concept showing that biotinylation proteomics of biofilms complemented with ex vivo proteomics of granuloma biofilms is a valuable strategy to uncover optimal nanobody targets within the biofilm matrix. Nanobody-based strategies have great potential to provide new and much-needed solutions for more effective treatment of tuberculosis, as well asother chronic biofilm infections with increased antibiotic tolerance and high risk for emerging antibiotic resistance
2023-03-30 | PXD033425 | Pride
Project description:The opportunistic pathogenic mold Aspergillus fumigatus is an increasing cause of morbidity and mortality in immunocompromised and, in part, immunocompetent patients. Like bacteria or yeast, A. fumigatus can grow in multicellular communities by the formation of a hyphal network encased in an extracellular matrix. Here, we describe the proteome and transcriptome of planktonic and biofilm-grown A. fumigatus mycelium after 24h and 48h. A biofilm- and time-dependent regulation of many proteins and genes of the primary metabolism indicates a developmental stage of the young biofilm at 24h, which demands energy. At a matured biofilm phase, metabolic activity seems to be reduced. However, genes encoding hydrophobins and proteins involved in the biosynthesis of secondary metabolites were significantly upregulated. In particular, proteins of the gliotoxin secondary metabolite gene cluster were induced in biofilm cultures. This was confirmed by RT-PCR and by detection of this immunologically active mycotoxin in culture supernatants using HPLC analysis. The enhanced production of gliotoxin by in vitro formed biofilms reported here may play also a significant role under in vivo conditions. It may confer A. fumigatus protection from the host immune system and also enable its survival and persistence in chronic lung infections such as aspergilloma.
2010-07-01 | GSE19430 | GEO
Project description:Biofilms with immobilized cells in industrial fermentation are beneficial. Encased in extracellular polymeric substance, cells forming biofilms are regulated by various factors. Nitric oxide (NO), as a signaling molecule, recognized as quorum sensing molecule regulating microbe biofilm formation. Regulation mechanisms of NO on bacteria biofilm have been studied extensively and deeply, while on fungus are rarely studied. In this study, we observed that low concentration of NO enhanced S. cerevisiae biofilm formation. Transcriptional and proteomic analysis revealed that transcription factor MAC1 was activated in biofilm cells under NO treatment. Overexpressed MAC1 increased yeast biofilm formation bypassing regulating the expression level of FLO11. Increased copper and iron contents in NO treated and MAC1 overexpressed cells were not responsible for increased biofilm formation. Among six downstream genes of MAC1, overexpressed CTR1 contributed yeast biofilm formation. Moreover, MAC1 and CTR1 contributed to biofilm cells ethanol resistance resulting from enhanced biofilm. The role of CTR1 protein in yeast biofilm formation may result from its hydrophobic residues in N-terminal extracellular domain. These findings suggested a NO-mediated biofilm formation mechanism that NO regulated expression levels of CTR1 through activating its transcription factor MAC1, leading enhanced biofilm formation.
2019-11-12 | PXD010751 | Pride