Regulation of amino sugar utilization in <scp><i>B</i></scp><i>acillus subtilis</i> by the <scp>GntR</scp> family regulators, <scp>NagR and GamR</scp>

Gaugué, Isabelle; Oberto, Jacques; Plumbridge, Jacqueline

doi:10.1111/mmi.12544

Cited by 23 publications

(34 citation statements)

References 37 publications

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“…The accumulation of DasR-6His into the wells was also observed at high concentrations of the pure protein without any tested ligand added in the EMSA reaction mix (Fig. S1C), a phenomenon that was also previously reported for the NagR protein of B. subtilis [18].…”

Section: Multiple Effects Of Phosphorylated (Amino)-sugars On the Dnasupporting

confidence: 58%

“…The first attempt to identify the allosteric effector of NagR revealed a similar inhibitory effect of GlcN-6P on the DNA-binding ability of NagR as described for DasR sco [17]. Later on, footprinting assays and EMSAs performed by Gaug e and colleagues did not show any effect of GlcN-6P and a very minor effect of GlcNAc-6P on the DNA-binding ability of NagR [18]. In contrast, the crystal structures of NagR in complex with GlcN-6-P and GlcNAc-6-P suggests that both molecules similarly displace the NagR-DNAbinding domains and therefore should similarly affect the DNAbinding ability of the regulator [19].…”

Section: Introductionmentioning

confidence: 99%

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Multiple allosteric effectors control the affinity of DasR for its target sites

Tenconi

Urem

Świątek-Połatyńska

et al. 2015

Biochemical and Biophysical Research Communications

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a b s t r a c tThe global transcriptional regulator DasR connects N-acetylglucosamine (GlcNAc) utilization to the onset of morphological and chemical differentiation in the model actinomycete Streptomyces coelicolor. Previous work revealed that glucosamine-6-phosphate (GlcN-6P) acts as an allosteric effector which disables binding by DasR to its operator sites (called dre, for DasR responsive element) and allows derepression of DasR-controlled/GlcNAc-dependent genes. To unveil the mechanism by which DasR controls S. coelicolor development, we performed a series of electromobility shift assays with histidinetagged DasR protein, which suggested that N-acetylglucosamine-6-phosphate (GlcNAc-6P) could also inhibit the formation of DasR-dre complexes and perhaps even more efficiently than GlcN-6P. The possibility that GlcNAc-6P is indeed an efficient allosteric effector of DasR was further confirmed by the high and constitutive activity of the DasR-repressed nagKA promoter in the nagA mutant, which lacks GlcNAc-6P deaminase activity and therefore accumulates GlcNAc-6P. In addition, we also observed that high concentrations of organic or inorganic phosphate enhanced binding of DasR to its recognition site, suggesting that the metabolic status of the cell could determine the selectivity of DasR in vivo, and hence its effect on the expression of its regulon.

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Section: Multiple Effects Of Phosphorylated (Amino)-sugars On the Dnasupporting

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Multiple allosteric effectors control the affinity of DasR for its target sites

Tenconi

Urem

Świątek-Połatyńska

et al. 2015

Biochemical and Biophysical Research Communications

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“…5B). We also noted clear patterns of mobility shifting for both probes, indicating that GlcN-6-P reduces the intermolecular interactions that are characteristic of NagR proteins (17) and that lead to aggregation of the DNA-protein complexes at the origin of the gel. Such dissociation would allow for the interactions between NagR and two dre sites to manifest as two distinct bands, with the upper band resulting from both dre sites being occupied (Fig.…”

Section: Fig 4 Cat Specific Activities (Expressed In Nanomoles Per MImentioning

confidence: 88%

“…In the Gram-positive bacterium Bacillus subtilis, a GntR/HutC-type regulator, designated NagR (formerly YvoA), binds to nonhomologous operator sites, also called dre (14,15), to control the transcription of nagA, nagB, and nagP, which codes for a GlcNAc-specific PTS permease. Unlike in E. coli, though, the allosteric effector(s) that alleviates repression of these target genes by NagR remains undetermined (14,16,17). Recently, an additional system encoded by ybgBA-gamAP and the GntR-type regulator GamR were discovered in B. subtilis and shown to encode products that regulate and effect the catabolism of GlcN (7,17).…”

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confidence: 99%

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NagR Differentially Regulates the Expression of the glmS and nagAB Genes Required for Amino Sugar Metabolism by Streptococcus mutans

Zeng

Burne

2015

J Bacteriol

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The ability of bacteria to metabolize glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) is considered important for persistent colonization of the oral cavity. In the dental caries pathogen Streptococcus mutans, the NagR protein regulates the expression of glmS, which encodes a GlcN-6-P synthetase, and nagA (GlcNAc-6-P deacetylase) and nagB (GlcN-6-P deaminase), which are required for the catabolism of GlcNAc and GlcN. Two NagR-binding sites (dre) were identified in each of the promoter regions for nagB and glmS. Using promoter-reporter gene fusions, the role of each dre site was examined in the regulation of glmS and nagB promoter activities in cells grown with glucose, GlcNAc, or GlcN. A synergistic relationship between the two dre sites in the glmS promoter that required proper spacing was observed, but that was not the case for nagB. Binding of purified NagR to DNA fragments from both promoter regions, as well as to dre sites alone, was strongly influenced by particular sugar phosphates. Using a random mutagenesis approach that targeted the effector-binding domain of NagR, mutants that displayed aberrant regulation of both the glmS and nagAB genes were identified. Collectively, these findings provide evidence that NagR is essential for regulation of genes for both the synthesis and catabolism of GlcN and GlcNAc in S. mutans, and that NagR engages differently with the target promoter regions in response to specific metabolites interacting with the effector-binding domain of NagR. IMPORTANCEGlucosamine and N-acetylglucosamine are among the most abundant naturally occurring sugars on the planet, and they are catabolized by many bacterial species as sources of carbon and nitrogen. Representing a group called lactic acid bacteria (LAB), the human dental caries pathogen Streptococcus mutans is shown to differ from known paradigm organisms in that it possesses a GntR/HutC-type regulator, NagR, that is required for the regulation of both catabolism of GlcN and biosynthesis. Results reported here reveal a simple and elegant mechanism whereby NagR differentially regulates two opposing biological processes by surveying metabolic intermediates. This study provides insights that may contribute to the development of novel therapeutic tools to combat dental caries and other infectious diseases.A mino sugars, including glucosamine (GlcN) and N-acetyl-Dglucosamine (GlcNAc or NAG), are some of the most abundant carbohydrates on earth. Since amino sugars can serve as sources of carbon and nitrogen, many bacteria have the capacity to utilize them for energy production and growth. We recently showed that the human dental pathogen Streptococcus mutans internalizes GlcN and GlcNAc via the phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS), which consists of a general enzyme I (EI) and a phospho-carrier protein (HPr) that transfer the phosphoryl group from PEP to a variety of substratespecific EII enzymes responsible for the concurrent phosphorylation and internalization of their cognate sugars (1). S. mu...

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Two novel regulators of N‐acetyl‐galactosamine utilization pathway and distinct roles in bacterial infections

Zhang

Ravcheev

et al. 2015

MicrobiologyOpen

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Bacterial pathogens can exploit metabolic pathways to facilitate their successful infection cycles, but little is known about roles of d‐galactosamine (GalN)/N‐acetyl‐d‐galactosamine (GalNAc) catabolism pathway in bacterial pathogenesis. Here, we report the genomic reconstruction of GalN/GalNAc utilization pathway in Streptococci and the diversified aga regulons. We delineated two new paralogous AgaR regulators for the GalN/GalNAc catabolism pathway. The electrophoretic mobility shift assays experiment demonstrated that AgaR2 (AgaR1) binds the predicted palindromes, and the combined in vivo data from reverse transcription quantitative polymerase chain reaction and RNA‐seq suggested that AgaR2 (not AgaR1) can effectively repress the transcription of the target genes. Removal of agaR2 (not agaR1) from Streptococcus suis 05ZYH33 augments significantly the abilities of both adherence to Hep‐2 cells and anti‐phagocytosis against RAW264.7 macrophage. As anticipated, the dysfunction in AgaR2‐mediated regulation of S. suis impairs its pathogenicity in experimental models of both mice and piglets. Our finding discovered two novel regulators specific for GalN/GalNAc catabolism and assigned them distinct roles into bacterial infections. To the best of our knowledge, it might represent a first paradigm that links the GalN/GalNAc catabolism pathway to bacterial pathogenesis.

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Regulation of amino sugar utilization in Bacillus subtilis by the GntR family regulators, NagR and GamR

Cited by 23 publications

References 37 publications

Multiple allosteric effectors control the affinity of DasR for its target sites

Multiple allosteric effectors control the affinity of DasR for its target sites

NagR Differentially Regulates the Expression of the glmS and nagAB Genes Required for Amino Sugar Metabolism by Streptococcus mutans

Two novel regulators of N‐acetyl‐galactosamine utilization pathway and distinct roles in bacterial infections

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