Regulation of Bacillus subtilis glutamine synthetase gene expression by the product of the glnR gene

Schreier, Harold J.; Brown, Stuart; Hirschi, Kendal D.; Nomellini, John F.; Sonenshein, Abraham L.

doi:10.1016/0022-2836(89)90290-8

Journal of Molecular Biology

1989

DOI: 10.1016/0022-2836(89)90290-8

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Regulation of Bacillus subtilis glutamine synthetase gene expression by the product of the glnR gene

Harold J. Schreier

Stuart Brown

Kendal D. Hirschi

et al.

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Cited by 120 publications

(176 citation statements)

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“…In the low-GC Gram-positive bacterium Bacillus subtilis, the GlnR and TnrA transcription factors regulate gene expression in response to nitrogen availability (1)(2)(3)(4). Both of these proteins are members of the MerR family of transcriptional regulators (2,4).…”

mentioning

confidence: 99%

“…Both of these proteins are members of the MerR family of transcriptional regulators (2,4). The sequences of the amino-terminal DNA-binding domains of these two proteins are highly similar, and they have identical DNA-binding site consensus sequences (4)(5)(6).…”

mentioning

confidence: 99%

“…The sequences of the amino-terminal DNA-binding domains of these two proteins are highly similar, and they have identical DNA-binding site consensus sequences (4)(5)(6). GlnR and TnrA have little sequence similarity in their carboxyl-terminal signaltransduction domains, and they are active under different nutritional conditions (2,4). GlnR represses gene expression during growth with excess nitrogen, whereas TnrA regulates transcription during nitrogen-limited growth.…”

mentioning

confidence: 99%

“…The expression of the B. subtilis glnRA operon, which contains the genes encoding GlnR (glnR) and GS (glnA), is repressed during growth with excess nitrogen (2). This dicistronic operon is found within the genomes of many low-GC Gram-positive bacteria (14).…”

mentioning

confidence: 99%

“…This dicistronic operon is found within the genomes of many low-GC Gram-positive bacteria (14). Genetic studies indicate that both GlnR and GS are required for the repression of glnRA expression (2,9,15,16).…”

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confidence: 99%

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Bacillus subtilis glutamine synthetase regulates its own synthesis by acting as a chaperone to stabilize GlnR–DNA complexes

Fisher

Wray

2008

Proc. Natl. Acad. Sci. U.S.A.

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The Bacillus subtilis GlnR repressor controls gene expression in response to nitrogen availability. Because all GlnR-regulated genes are expressed constitutively in mutants lacking glutamine synthetase (GS), GS is required for repression by GlnR. Feedbackinhibited GS (FBI-GS) was shown to activate GlnR DNA binding with an in vitro electophoretic mobility shift assay (EMSA). The activation of GlnR DNA binding by GS in these experiments depended on the feedback inhibitor glutamine and did not occur with mutant GS proteins defective in regulating GlnR activity in vivo. Although stable GS-GlnR-DNA ternary complexes were not observed in the EMSA experiments, cross-linking experiments showed that a protein-protein interaction occurs between GlnR and FBI-GS. This interaction was reduced in the absence of the feedback inhibitor glutamine and with mutant GS proteins. Because FBI-GS significantly reduced the dissociation rate of the GlnR-DNA complexes, the stability of these complexes is enhanced by FBI-GS. These results argue that FBI-GS acts as a chaperone that activates GlnR DNA binding through a transient protein-protein interaction that stabilizes GlnR-DNA complexes. GS was shown to control the activity of the B. subtilis nitrogen transcription factor TnrA by forming a stable complex between FBI-GS and TnrA that inhibits TnrA DNA binding. Thus, B. subtilis GS is an enzyme with dual catalytic and regulatory functions that uses distinct mechanisms to control the activity of two different transcription factors.nitrogen regulation ͉ MerR ͉ TurA

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confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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Bacillus subtilis glutamine synthetase regulates its own synthesis by acting as a chaperone to stabilize GlnR–DNA complexes

Fisher

Wray

2008

Proc. Natl. Acad. Sci. U.S.A.

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Synergetic engineering of central carbon and nitrogen metabolism for the production of N‐acetylglucosamine in Bacillus subtilis

Niu

Liu²,

et al. 2019

Biotech and App Biochem

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N‐acetylglucosamine (GlcNAc) is a nitrogen‐containing compound, which is widely used as a nutraceutical and pharmaceutical. In our previous work, we constructed a recombinant Bacillus subtilis strain for the biosynthesis of GlcNAc by engineering the central carbon metabolism. However, nitrogen is also required for the synthesis of GlcNAc. Hence, it is necessary to simultaneously coordinate the carbon and nitrogen metabolism to improve production of GlcNAc. In this work, we attempted to enhance GlcNAc production in B. subtilis by increasing supply of precursors N‐acetylglucosamine 6‐phosphate (GlcNAc6P) and glutamate. The expression of a key enzyme, GlcNAc6P N‐acetyltransferase (GNA1), was enhanced by engineering the promoter and ribosome binding site to enhance the production of GlcNAc6P. Next, we examined the effect of different nitrogen sources on GlcNAc synthesis. We observed that urea can promote nitrogen assimilation for GlcNAc synthesis. The glutamate synthesis was improved by deleting the two endogenous glutamate dehydrogenase genes (rocG and gudB) and by integrating one exogenous glutamate dehydrogenase gene (gdh). This strategy enhanced the intracellular glutamate and glutamine by 69.8% and 46.9%, respectively. The synergetic engineering of central carbon and nitrogen metabolisms increased the GlcNAc titer from 14.0 to 22.2 g/L in the shaker flask. Hence, our study demonstrated the importance of carbon and nitrogen metabolism coordination in the production of nitrogen‐containing compounds.

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Global expression profiling of Bacillus subtilis cells during industrial‐close fed‐batch fermentations with different nitrogen sources

Brinckmann

Tobisch

Wümpelmann

et al. 2005

Biotech & Bioengineering

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A detailed gene expression analysis of industrial-close Bacillus subtilis fed-batch fermentation processes with casamino acids as the only nitrogen source and with a reduced casamino acid concentration but supplemented by ammonia was carried out. Although glutamine and arginine are supposed to be the preferred nitrogen sources of B. subtilis, we demonstrate that a combined feeding of ammonia and casamino acids supports cell growth under fed-batch fermentation conditions. The transcriptome and proteome analyses revealed that the additional feeding of ammonia in combination with a reduced amino acid concentration results in a significantly lower expression level of the glnAR or tnrA genes, coding for proteins, which are mainly involved in the nitrogen metabolism of B. subtilis. However, the mRNA levels of the genes of the ilvBHC-leuABD and hom-thrCB operons were significantly increased, indicating a valine, leucine, isoleucine, and threonine limitation under these fermentation conditions. In contrast, during the fermentation with casamino acids as the only nitrogen source, several genes, which play a crucial role in nitrogen metabolism of B. subtilis (e.g., glnAR, nasCDE, nrgAB, and ureABC), were up-regulated, indicating a nitrogen limitation under these conditions. Furthermore, increased expression of genes, which are involved in motility and chemotaxis (e.g., hag, fliT) and in acetoin metabolism (e.g., acoABCL), was determined during the fermentation with the mixed nitrogen source of casamino acids and ammonia, indicating a carbon limitation under these fermentation conditions. Under high cell density and slow growth rate conditions a weak up-regulation of autolysis genes could be observed as well as the induction of a number of genes involved in motility, chemotaxis and general stress response. Results of this study allowed the selection of marker genes, which could be used for the monitoring of B. subtilis fermentation processes. The data suggest for example acoA as a marker gene for glucose limitation or glnA as an indicator for nitrogen limitation.

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Regulation of Bacillus subtilis glutamine synthetase gene expression by the product of the glnR gene

Cited by 120 publications

References 30 publications

Bacillus subtilis glutamine synthetase regulates its own synthesis by acting as a chaperone to stabilize GlnR–DNA complexes

Bacillus subtilis glutamine synthetase regulates its own synthesis by acting as a chaperone to stabilize GlnR–DNA complexes

Synergetic engineering of central carbon and nitrogen metabolism for the production of N‐acetylglucosamine in Bacillus subtilis

Global expression profiling of Bacillus subtilis cells during industrial‐close fed‐batch fermentations with different nitrogen sources

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