Exploring roles of the chitinase ChiC in modulating Pseudomonas aeruginosa virulence phenotypes
Ontology highlight
ABSTRACT:
INSTRUMENT(S): timsTOF Pro
ORGANISM(S): Pseudomonas Aeruginosa Pao1 Bacteria
SUBMITTER: 
Per Kristian Edvardsen
LAB HEAD: Gustav Vaaje-Kolstad
PROVIDER: PXD048596 | Pride | 2024-04-30
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| 210826_KO_ctrl_1.d.zip | Other | |||
| 210826_KO_ctrl_2.d.zip | Other | |||
| 210826_KO_ctrl_3.d.zip | Other | |||
| 210826_KO_pig_mucin_1.d.zip | Other | |||
| 210826_KO_pig_mucin_2.d.zip | Other |
Items per page: 5 1 - 5 of 14 |
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Project description:Chitinases are ubiquitous enzymes involved in biomass degradation and chitin turnover in nature. Pseudomonas aeruginosa (PA), an opportunistic human pathogen, expresses ChiC, a secreted glycoside hydrolase 18 (GH18) family chitinase. Despite speculation about ChiC's role in PA disease pathogenesis, there is scant evidence supporting this hypothesis. Since PA cannot catabolize chitin, we investigated the potential function(s) of ChiC in PA pathophysiology
2024-04-30 | PXD048320 | Pride
Project description:Understanding the strategies used by bacteria to degrade polysaccharides constitutes an invaluable tool for biotechnological applications Bacteria are major mediators of polysaccharide degradation in nature, however the complex mechanisms used to detect, degrade, and consume these substrates are not well understood, especially for recalcitrant polysaccharides such as chitin It has been previously shown that the model bacterial saprophyte Cellvibrio japonicus is able to catabolize chitin, but little is known about the enzymatic machinery underlying this capability Previous analyses of the C japonicus genome and proteome indicated the presence of four family 18 Glycoside Hydrolase (GH18) enzymes, and studies of the proteome indicated that all are involved in chitin utilization Using a combination of in vitro and in vivo approaches, we have studied the roles of these four chitinases in chitin bioconversion Genetic analyses showed that only the chi18D gene product is essential for the degradation of chitin substrates Biochemical characterization of the four enzymes showed functional differences and synergistic effects during chitin degradation, indicating non-redundant roles in the cell Transcriptomic studies revealed complex regulation of the chitin degradation machinery of C japonicus and confirmed the importance of CjChi18D and CjLPMO10A, a previously characterized chitin-active enzyme With this systems biology approach, we deciphered the physiological relevance of the GH18 enzymes for chitin degradation in C japonicus, and the combination of in vitro and in vivo approaches provided a comprehensive understanding of the initial stages of chitin degradation by this bacterium
2018-01-24 | GSE108935 | GEO
Project description:Chitin is an abundant natural polysaccharide that is hard to degrade because of its crystalline nature and because it is often found embedded in robust co-polymeric materials containing other polysaccharides, proteins and minerals. Thus, it is of interest to study the enzymatic machineries of highly specialized microbes found in chitin-rich environments. We describe a genomic and proteomic analysis of the chitinolytic machinery of Andreprevotia ripae, a Gram-negative bacterium that was isolated from an anthill. The genome of A. ripae encodes four secreted family GH19 chitinases of which two were detected, both being upregulated during growth on chitin. In addition, the genome encodes 25 secreted GH18 chitinases, of which 17 were detected and 12 were upregulated during growth on chitin. Finally, the single lytic polysaccharide monooxygenase (LPMO) of A. ripae, predicted to be chitin-active based on sequence characteristics and the presence of chitin-binding domains, was strongly upregulated during growth on chitin. Whereas 66 % of the 29 secreted chitinases contained two carbo¬hydrate-binding modules (CBMs) known for binding to chitin, this fraction was 93 % (13 out of 14) for the chitinases that were upregulated during growth on chitin, suggesting an important role for these CBMs. Next to demonstrating an unprecedentedly large multiplicity of upregulated chitinases, the present study also revealed several chitin-induced proteins that contain chitin-binding domains but lack a known catalytic function. These proteins are interesting targets for discovery of enzymes used by Nature to convert chitin-rich biomass.
2021-06-10 | PXD025087 | Pride
Project description:The marine bacterium Aliivibriosalmonicida (formerly Vibrio salmonicida) is the causative agent of cold water vibriosis, a disease that led to severe losses in Norwegian aquaculture during the 1980s. Genes encoding a glycoside hydrolase family 18 (GH18) chitinase and two lytic polysaccharide monooxygenases (LPMOs) containing chitin binding domains are encoded in the genome of this bacterium, indicating a chitin degrading capability. However, due to extensive gene decay, some parts of the chitinolytic pathways are believed to be dysfunctional. To investigate the chitinolytic abilities of the bacterium, we combined microbiological and biochemical experiments with proteomic analysis. The family GH18 chitinase was able to depolymerize - and -chitin, but its activity was up to 50-fold lower compared to other well-studied chitinases from Serratia marcescens and Cellvibrio japonicus. The two LPMOs showed activity on chitin, cleaving the substrate chains by oxidation. Cultivation experiments showed that Al. salmonicida was able to grow on and utilize the soluble building blocks of chitin; N-acetyl-D-glucosamine (GlcNAc) and chitobiose (GlcNAc2). Furthermore, cultivation and gene deletion studies revealed that the bacterium was able to degrade insoluble chitin, albeit at a slow rate, and that growth on this substrate was dependent on the GH18 chitinase, but to a lesser extent the LPMOs. Finally, expression of the GH18 chitinase and both LPMOs was detected by proteomic analysis of Al. salmonicida cultivated on chitin as the sole carbon source. In conclusion, our results show that Al. salmonicida LFI1238 can utilize chitin as source of nutrients and its ability depends on the GH18 chitinase.
2021-07-26 | PXD021397 | Pride
Project description:Genome mining of pigmented Pseudoalteromonas has revealed a large potential for production of bioactive compounds, both hydrolytic enzymes and secondary metabolites and the purpose of the present study was to explore this bioactivity potential in a potent antibiotic and enzyme producer, Pseudoalteromonas rubra strain S4059. Proteomic analyses indicated that a highly efficient chitin degradation machinery was present in the red-pigmented P. rubra S4059 when grown on chitin. Four GH18 chitinases and two GH20 hexosaminidases were significantly upregulated by chitin. GH19 chitinase which is not common in bacteria is consistently found in pigmented Pseudoalteromonas and in S4059 it was only detected when the bacterium was grown on chitin. To explore the possible role of GH19 in pigmented Pseudoalteromonas, we deleted the GH19 chitinase and compared a range of phenotypes in the mutant and wild type. Neither, the chitin degrading ability or the biofilm forming capacity was affected by GH19 deletion. In some Vibrionaceae, the secondary metabolome is significantly affected by growth on chitin as compared to simpler carbon sources. The secondary metabolites produced by S4059 and the GH19 mutant were xxx start by chitin/mannose – then the mutant. not altered by the absence of the gene, indicating that chitin utilization may not directly influence the production of secondary metabolites as has been observed in some Vibrionaceae. Metabolome analysis reveal that growth on chitin XX. In summary,
2021-03-11 | PXD023249 | Pride
Project description:The Atlantic salmon (Salmo salar) genome contains 10 chitinase encoding genes, but little is known about the function of these chitinases. Three of the chitinase genes have previously been shown to be expressed in the stomach tissue of Atlantic salmon. In the current study we show that the protein products of these genes, the family 18 glycoside hydrolase (GH18) chitinases, Chia.3, Chia.4 and Chia.7 are secreted into the stomach mucosa and are amongst the most abundant proteins in this matrix.
2022-05-08 | PXD030291 | Pride
Project description:This SuperSeries is composed of the following subset Series: GSE3101: Chitin oligosaccharide induction GSE3102: Crab shell attachment GSE3103: Chitin sensor Abstract: Chitin, an insoluble polymer of GlcNAc, is an abundant source of carbon, nitrogen, and energy for marine microorganisms. Microarray expression profiling and mutational studies of Vibrio cholerae growing on a natural chitin surface, or with the soluble chitin oligosaccharides (GlcNAc)(2-6), GlcNAc, or the glucosamine dimer (GlcN)2 identified three sets of differentially regulated genes. We show that (i) ChiS, a sensor histidine kinase, regulates expression of the (GlcNAc)(2-6) gene set, including a (GlcNAc)2 catabolic operon, two extracellular chitinases, a chitoporin, and a PilA-containing type IV pilus, designated ChiRP (chitin-regulated pilus) that confers a significant growth advantage to V. cholerae on a chitin surface; (ii) GlcNAc causes the coordinate expression of genes involved with chitin chemotaxis and adherence and with the transport and assimilation of GlcNAc; (iii) (GlcN)2 induces genes required for the transport and catabolism of nonacetylated chitin residues; and (iv) the constitutively expressed MSHA pilus facilitates adhesion to the chitin surface independent of surface chemistry. Collectively, these results provide a global portrait of a complex, multistage V. cholerae program for the efficient utilization of chitin. Refer to individual Series
2006-03-07 | E-GEOD-4388 | biostudies-arrayexpress
Project description:Abstract: Chitin, an insoluble polymer of GlcNAc, is an abundant source of carbon, nitrogen, and energy for marine microorganisms. Microarray expression profiling and mutational studies of Vibrio cholerae growing on a natural chitin surface, or with the soluble chitin oligosaccharides (GlcNAc)(2-6), GlcNAc, or the glucosamine dimer (GlcN)2 identified three sets of differentially regulated genes. We show that (i) ChiS, a sensor histidine kinase, regulates expression of the (GlcNAc)(2-6) gene set, including a (GlcNAc)2 catabolic operon, two extracellular chitinases, a chitoporin, and a PilA-containing type IV pilus, designated ChiRP (chitin-regulated pilus) that confers a significant growth advantage to V. cholerae on a chitin surface; (ii) GlcNAc causes the coordinate expression of genes involved with chitin chemotaxis and adherence and with the transport and assimilation of GlcNAc; (iii) (GlcN)2 induces genes required for the transport and catabolism of nonacetylated chitin residues; and (iv) the constitutively expressed MSHA pilus facilitates adhesion to the chitin surface independent of surface chemistry. Collectively, these results provide a global portrait of a complex, multistage V. cholerae program for the efficient utilization of chitin. This SuperSeries is composed of the SubSeries listed below.
2006-03-08 | GSE4388 | GEO
Project description:Purpose: Evaluate the anti-tumorigenic efficacy and immunomodulatory effects of chitin as general blocker of immunosuppressive chitinase-like proteins (CLPs) in combination with and without anti-PD-1 immune checkpoint blockade (ICB) in an immunocompetent 4T1- and 66cl4-based intraductal model for triple-negative breast cancer (TNBC). Methods: 4T1 and 66cl4 mammary tumor-bearing female BALB/c mice were either left untreated or were treated with anti-PD-1, chitin or chitin + anti-PD-1 for 2 weeks (w) between 3 and 5 w post-inoculation (p.i.) of the mammary tumor cells. After the 2-w treatment, 4T1 and 66cl4 primary tumors were resected and RNA was isolated from the tissue using in-house developed protocols. Results: Chitin-mediated reduction in innate immunosuppression and increased anti-tumor T-cell immunity in primary tumors of both TNBC models was clearly presented at the genomic level based on RNA-sequencing analysis. More specifically, a selection of 68 and 55 innate immunity-related genes in respectively 4T1 and 66cl4 primary tumors were downregulated upon chitin treatment in combination with or without anti-PD-1. Several of the selected innate immunity genes could be linked to immunosuppressive myeloid cell types, including Ccl2, Cxcl2, Csf1r and Itgam. Genes related to T-cell exhaustion, including Pdcd1, Cd274, Havcr2 and Lag3, were downregulated upon chitin and chitin + anti-PD-1 treatment in 4T1 tumors. Chitin- and chitin + anti-PD-1-treated 66cl4 tumors showed upregulation of genes associated with enhanced T-cell activity, including Cd8a, Cd8b1, Ifng, Il12b and Il18r1. In line with an overall reduced inflammation upon chitin treatment at the protein level, predominantly through myeloid cell reduction, a selected set of 68 and 48 inflammation-related genes were downregulated in chitin- and chitin + anti-PD-1-treated primary tumors of respectively the 4T1- and the 66cl4-based model. Importantly, genes that have been associated with the signaling and pro-tumorigenic activity of a prominent CLP family member, i.e. CHI3L1, were also downregulated in primary tumors following chitin treatment either with or without anti-PD-1, including Usf1 and Rab37 in the 4T1-based model and Lgals3 in the 66cl4-based model. Conclusion: Our findings highlight that chitin, as a general CLP blocker, reduces cancer-associated immunosuppression and enhances anti-tumor immunity as well as ICB responses in complementary ICB-resistant TNBC models, supporting its potential clinical relevance in immunosuppressed TNBC patients.
2024-02-25 | GSE256439 | GEO
Project description:Chitin utilization by microbes plays a significant role in cycling of carbon and nitrogen in the biosphere, and the study of the microbial approaches used to degrade chitin will facilitate our understanding of bacterial strategies to degrade this recalcitrant polysaccharide. The early stages of chitin depolymerization by the bacterium Cellvibrio japonicus have been characterized and are dependent on one chitin-specific lytic polysaccharide monooxygenase and non-redundant glycoside hydrolases from the family GH18 to generate chito-oligosaccharides for entry into metabolism. Here, we describe the mechanisms for the latter stages of chitin utilization by C. japonicus with an emphasis on the fate of chito-oligosaccharides. Our systems biology approach combined transcriptomics, bacterial genetics, and complex environmental substrates to determine the essential mechanisms for chito-oligosaccharide transport and catabolism in Cellvibrio japonicus. Using RNAseq analysis we found not only the up-regulation of genes that encode CAZymes specific for chitin metabolism but also a coordinated expression of non-chitin specific CAZyme genes. Furthermore, we used mutational analysis to characterize the hex20B gene product, predicted to encode a hexosaminidase, and found that it is required for efficient utilization of chito-oligosaccharides. Surprisingly, two additional gene loci (CJA_0353 and CJA_1157), which encode putative TonB-dependent transporters, were essential for shuttling chito-oligosaccharides into the periplasmic space. Here we propose naming these loci cttA (chito-oligosaccharides transporter A) and cttB respectively. This study further develops our model of how C. japonicus can derive nutrients from recalcitrant chitin-containing substrates and may be potentially useful for other environmentally or industrially important bacteria.
2020-04-30 | GSE149593 | GEO