Abstract
NagR, belonging to the GntR/HutC family, is a negative regulator that directly represses the nagP and nagAB genes, which are involved in GlcNAc transport and utilization in Bacillus subtilis. Our previous work confirmed that the chitinase B gene (chiB) of Bacillus thuringiensis strain Bti75 is also negatively controlled by YvoABt, the ortholog of NagR from B. subtilis. In this work, we investigated its regulatory network in Bti75 and found that YvoABt is an N-acetylglucosamine utilization regulator primarily involved in GlcNAc catabolism; therefore YvoABt is renamed as NagRBt. The RNA-seq data revealed that 27 genes were upregulated and 14 genes were downregulated in the ΔnagR mutant compared with the wild-type strain. The regulon (exponential phase) was characterized by RNA-seq, bioinformatics software, electrophoretic mobility shift assays, and quantitative real-time reverse transcription PCR. In the Bti75 genome, 19 genes that were directly regulated and 30 genes that were indirectly regulated by NagRBt were identified. We compiled in silico, in vitro, and in vivo evidence that NagRBt behaves as a repressor and activator to directly or indirectly influence major biological processes involved in amino sugar metabolism, nucleotide metabolism, fatty acid metabolism, phosphotransferase system, and the Embden–Meyerhof–Parnas pathway.
Highlights
N-Acetylglucosamine (GlcNAc) is a nitrogen-containing monosaccharide that is a preferred nutrient source for the growth and development of many microorganisms because it is highly abundant and provides both carbon and nitrogen (Mobley et al, 1982)
Later studies revealed that DasR is a global regulator that plays a pivotal role in the regulation of antibiotic synthesis, morphological differentiation, and GlcNAc transport and NagRBt Is a Pleiotropic and Dual Transcriptional Regulator metabolism (Rigali et al, 2006, 2008; Swiatek et al, 2012; Tenconi et al, 2015)
We demonstrate that NagRBt acts as a pleiotropic transcriptional regulator that controls at least 19 genes directly and 30 genes indirectly, and these genes are involved in amino sugar metabolism, nucleotide metabolism, fatty acid metabolism, phosphotransferase system, and the Embden–Meyerhof–Parnas (EMP) pathway
Summary
N-Acetylglucosamine (GlcNAc) is a nitrogen-containing monosaccharide that is a preferred nutrient source for the growth and development of many microorganisms because it is highly abundant and provides both carbon and nitrogen (Mobley et al, 1982). GlcNAc is funneled into the glycolysis shunt pathway for catabolism or directed to peptidoglycan synthesis for anabolism (Bertram et al, 2011). GlcNAc is exploited for both catabolic and anabolic purposes; its correct utilization requires rigorous control. Rigali et al found the role of DasR in GlcNAc metabolism using Streptomyces coelicolor as a model organism (Rigali et al, 2004). Later studies revealed that DasR is a global regulator that plays a pivotal role in the regulation of antibiotic synthesis, morphological differentiation, and GlcNAc transport and NagRBt Is a Pleiotropic and Dual Transcriptional Regulator metabolism (Rigali et al, 2006, 2008; Swiatek et al, 2012; Tenconi et al, 2015). DasR can bind the dre sites in the upstream regions of some genes to control their expressions
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