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The ROK-family regulator RokL6 is involved in GlcN toxicity by controlling the transcription of SCO1448 in Streptomyces coelicolor [ChIP-seq]

ABSTRACT: Streptomycetes are saprophytic bacteria that grow on complex polysaccharides, such as cellulose, starch, chitin and chitosan. For the monomeric building blocks glucose, maltose and N-acetylglucosamine the metabolic pathway is well documented, but that of glucosamine (GlcN) is currently unknown. Importantly, GlcN is lethal to Streptomyces coelicolor nagB mutants, which lack glucosamine-6-phosphate deaminase activity. Here we report that spontaneous and directed mutations in the gene for the ROK-family protein RokL6 (SCO1447) relieve GlcN toxicity in nagB mutants of S. coelicolor. RNA sequencing, ChIP-Seq and over-expression studies revealed that RokL6 acts by directly repressing SCO1448, which encodes a sugar exporter; RokL6 thereby only binds to the rokL6-SCO1448 intergenic region in vivo, with consensus binding site C(T)TATCAGG - 7 nt - CCTGATAG(A). The exact transcriptional start sites for rokL6 and SCO1448 were determined using 5’RACE. RokL6 represses the transcription of both rokL6 and SCO1448 by binding to overlapping promoter sequences. Taken together, our data show that RokL6 and SCO1448 are novel GlcN-related genes, whereby RokL6 directly controls the transcription of SCO1448. The latter is a key protein in the defense of S. coelicolor against the toxicity of GlcN in a nagB-mutant background, most likely via the export of GlcN-derived toxic intermediates.

ORGANISM(S): Streptomyces coelicolor A3(2)

PROVIDER: GSE234438 | GEO | 2023/06/19

REPOSITORIES: GEO

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The ROK-family regulator RokL6 is involved in GlcN toxicity by controlling the transcription of SCO1448 in Streptomyces coelicolor [RNA-seq]

Project description:Streptomycetes are saprophytic bacteria that grow on complex polysaccharides, such as cellulose, starch, chitin and chitosan. For the monomeric building blocks glucose, maltose and N-acetylglucosamine the metabolic pathway is well documented, but that of glucosamine (GlcN) is currently unknown. Importantly, GlcN is lethal to Streptomyces coelicolor nagB mutants, which lack glucosamine-6-phosphate deaminase activity. Here we report that spontaneous and directed mutations in the gene for the ROK-family protein RokL6 (SCO1447) relieve GlcN toxicity in nagB mutants of S. coelicolor. RNA sequencing, ChIP-Seq and over-expression studies revealed that RokL6 acts by directly repressing SCO1448, which encodes a sugar exporter; RokL6 thereby only binds to the rokL6-SCO1448 intergenic region in vivo, with consensus binding site C(T)TATCAGG - 7 nt - CCTGATAG(A). The exact transcriptional start sites for rokL6 and SCO1448 were determined using 5’RACE. RokL6 represses the transcription of both rokL6 and SCO1448 by binding to overlapping promoter sequences. Taken together, our data show that RokL6 and SCO1448 are novel GlcN-related genes, whereby RokL6 directly controls the transcription of SCO1448. The latter is a key protein in the defense of S. coelicolor against the toxicity of GlcN in a nagB-mutant background, most likely via the export of GlcN-derived toxic intermediates.

2023-06-19 | GSE234437 | GEO

Expression profiles of dnaA null, rok null, and dnaA null; rok null mutants

2015-06-13 | GSE69788 | GEO

Expression profiles of dnaA null, rok null, and dnaA null; rok null mutants

Project description:To identify potential targets of co-regulation by DnaA and Rok, we compared the transcriptional profiles of dnaA null, rok null, and dnaA null rok null mutants. Because a dnaA null mutant requires an oriC- strain background, we used an oriC- oriN+ background for all strains, to allow direct comparisons. We compared biological triplicates for each of the following strains: dnaA null (CAL2074), rok null (CAS196), dnaA null rok null (CAS192), and dnaA+ rok+ (CAL2083; control).

2015-06-13 | E-GEOD-69788 | biostudies-arrayexpress

Role of Rok in biofilm structure development of Bacillus subtilis 168

Project description:Comparison of the B subtilis rok mutant vs wild type (sample 1-4) and rok-comK mutant vs comK mutant (sample 5-8)

2011-02-01 | GSE22370 | GEO

Role of Rok in biofilm structure development of Bacillus subtilis 168

Project description:Comparison of the B subtilis rok mutant vs wild type (sample 1-4) and rok-comK mutant vs comK mutant (sample 5-8) One condition design comparision of (rok vs wt) and (rok-comK vs comK) including a dye swap, 4 biological replicate

2011-02-01 | E-GEOD-22370 | biostudies-arrayexpress

Chromatin immunoprecipitation of Rok-myc and Hbs-myc in Bacillus subtilis

2011-01-01 | GSE23199 | GEO

Chromatin immunoprecipitation of Rok-myc and Hbs-myc in Bacillus subtilis

Project description:The Rok protein of Bacillus subtilis was identified as a negative regulator of competence development. Here we show that Rok binds to extended areas of the B. subtilis genome that are characterized by a high A+T content and are known or believed to have been acquired by horizontal gene transfer, e.g. mobile elements. A deletion of rok results in higher excision of one such element, ICEBs1. The preferential association of Rok with DNA with a high A+T content is also observed in a Gram-negative host, E. coli, and depends on a conserved C-terminal region of the protein. Based on our findings, we propose that Rok is a nucleoid-associated protein that fulfills a function analogous to H-NS, a protein absent from most Gram-positive bacteria. The genome-wide binding profile of the transcription factor Rok and the nucleoid-binding protein HBsu were determined. Three biological replicates were analyzed per strain (one per array). Binding profiles were determined in exponentially growing cells. Enrichment in immunoprecipitated samples versus total genomic DNA were determined.

2011-01-01 | E-GEOD-23199 | biostudies-arrayexpress

A unique cell wall synthetic response evoked by glucosamine determines pathogenicity-associated fungal cellular differentiation

Project description:The yeast-to-hypha transition is tightly associated with pathogenicity in many human pathogenic fungi, such as the model fungal pathogen Cryptococcus neoformans, which is responsible for approximately 180,000 deaths annually. In this pathogen, the yeast-to-hypha transition can be initiated by distinct stimuli: mating stimulation or glucosamine (GlcN), the monomer of cell wall chitosan. However, it remains poorly understood how the signal specificity for Cryptococcus morphological transition by disparate stimuli is ensured. Here, by integrating temporal expression signature analysis and phenome-based clustering evaluation, we demonstrate that GlcN specifically triggers a unique cellular response, which acts as a critical determinant underlying the activation of GlcN-induced filamentation (GIF). This cellular response is defined by an unusually hyperactive cell wall synthesis that is highly ATP-consuming. A novel cell surface protein Gis1 was identified as the indicator molecule for the GlcN-induced cell wall response. The Mpk1-directed cell wall pathway critically bridges global cell wall gene induction and intracellular ATP supply, ensuring the Gis1-dependent cell wall response and the stimulus specificity of GIF. We further reveal that the ability of Mpk1 to coordinate the cell wall response and GIF activation is conserved in different Cryptococcus pathogens. Phosphoproteomics-based profiling together with genetic and phenotypic analysis revealed that the Mpk1 kinase mediates the regulatory specificity of GIF through a coordinated downstream regulatory network centered on Skn7 and Crz1. Overall, our findings discover an unprecedented and conserved cell wall biosynthesis-dependent fungal differentiation commitment mechanism, which enables the signal specificity of pathogenicity-related dimorphism induced by GlcN in Cryptococcus pathogens.

2021-08-27 | GSE146435 | GEO

A unique cell wall synthetic response evoked by glucosamine determines pathogenicity-associated fungal cellular differentiation

2021-09-15 | PXD018015 | Pride

Expression data from glucosamine-treated human synovial MH7A cells

Project description:Osteoarthritis (OA) is one of the major joint diseases. Glucosamine (GlcN) is widely used as a dietary supplement for OA. However, its precise mechanisms are not well understood. GlcN is utilized for the O-linked-N-acetylglucosamine (O-GlcNAc) modification of proteins, which participates in the regulation of cellular functions. In this study, we comprehensively analyzed the effect of GlcN and alloxan (O-GlcNAc transferase inhibitor, which prevented O-GlcNAc modification) on the gene expression using DNA microarray. GlcN downregulated or upregulated genes. Furthermore, among the GlcN-downregulated or upregulated genes, the expression of 62.7% of the genes was restored by alloxan.. Thus, it is suggested that GlcN regulates the gene expression by an O-GlcNAc modification-dependent or -independent mechanism.

2016-11-07 | GSE72575 | GEO

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