CcpA Mutants with Differential Activities in &lt;i&gt;Bacillus subtilis&lt;/i&gt;

Sprehe, Mareen; Seidel, Gerald; Diel, Marco; Hillen, Wolfgang

doi:10.1159/000096464

Cited by 18 publications

(29 citation statements)

References 61 publications

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“…In contrast, those sites located upstream of the Ϫ35 promoter sequence are generally involved in activation (48). Recognition of these sites may also require CcpA to interact with Hpr-Ser-P, although CcpA may be in a different conformational state (42). In this regard, activation of the speB gene by CcpA presents an interesting example.…”

Section: Discussionmentioning

confidence: 99%

Distinct Time-Resolved Roles for Two Catabolite-Sensing Pathways during Streptococcus pyogenes Infection

Kietzman

Caparon

2011

Infect Immun

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Many Gram-positive pathogens link the expression of virulence genes to the presence of carbon source substrates using overlapping pathways for global control of carbon catabolite regulation. However, how these pathways are integrated to control the behavior of the transcriptome in time-and compartment-specific patterns is typically not well understood. In the present study, global transcriptome profiling was used to determine the extent to which glucose alters gene expression in Streptococcus pyogenes (group A streptococcus) and the contributions of the CcpA and LacD.1 catabolite control pathways to the regulation of this response in vitro. This analysis revealed that the expression of as many as 15% of the genes examined was regulated and that CcpA and LacD.1 together contribute to the regulation of 60% of this subset. However, numerous patterns were observed, including both CcpA-and LacD.1-specific and independent regulation, coregulation, and regulation of genes by these pathways independently of glucose. In addition, CcpA and LacD.1 had antagonistic effects on most coregulated genes. To resolve the roles of these regulators during infection, the expression of selected transcripts representative of different regulatory patterns was examined in a murine model of soft tissue infection. This revealed distinct patterns of misregulation with respect to time in CcpA ؊ versus LacD.1 ؊ mutants. Taken together, these data support an important role for carbohydrate in the regulation of the transcriptome in tissue and suggest that the CcpA and LacD.1 pathways are organized to function at different times during the course of an infection.

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Section: Discussionmentioning

confidence: 99%

Distinct Time-Resolved Roles for Two Catabolite-Sensing Pathways during Streptococcus pyogenes Infection

Kietzman

Caparon

2011

Infect Immun

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“…In Escherichia coli and Bacillus subtilis, regulators like Crp (cyclic AMP [cAMP] receptor protein) and CcpA (catabolite control protein A) not only regulate pts genes but also are global regulators of carbon metabolism in these bacteria. CcpA is the master regulator of carbon catabolite regulation in B. subtilis (30,53) and regulates more than 300 genes by either activation (e.g., the ␣-acetolactate synthase gene alsS and the acetate kinase gene ackA) or repression (e.g., the gluconate operon repressor gene gntR) (29,44,53). Activation and repression mediated by CcpA may utilize different conformational changes of the protein (53).…”

mentioning

confidence: 99%

“…CcpA is the master regulator of carbon catabolite regulation in B. subtilis (30,53) and regulates more than 300 genes by either activation (e.g., the ␣-acetolactate synthase gene alsS and the acetate kinase gene ackA) or repression (e.g., the gluconate operon repressor gene gntR) (29,44,53). Activation and repression mediated by CcpA may utilize different conformational changes of the protein (53). Because ccpA mutants are unable to activate glycolysis or carbon overflow metabolism, CcpA appears to control a superregulon of glucose catabolism in this organism (61).…”

mentioning

confidence: 99%

The Global Repressor SugR Controls Expression of Genes of Glycolysis and of the l -Lactate Dehydrogenase LdhA in Corynebacterium glutamicum

Engels

Lindner²,

Wendisch³

2008

J Bacteriol

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The transcriptional regulator SugR from Corynebacterium glutamicum represses genes of the phosphoenolpyruvate-dependent phosphotransferase system (PTS). Growth experiments revealed that the overexpression of sugR not only perturbed the growth of C. glutamicum on the PTS sugars glucose, fructose, and sucrose but also led to a significant growth inhibition on ribose, which is not taken up via the PTS. Chromatin immunoprecipitation combined with DNA microarray analysis and gel retardation experiments were performed to identify further target genes of SugR. Gel retardation analysis confirmed that SugR bound to the promoter regions of genes of the glycolytic enzymes 6-phosphofructokinase (pfkA), fructose-1,6-bisphosphate aldolase (fba), enolase (eno), pyruvate kinase (pyk), and NAD-dependent L-lactate dehydrogenase (ldhA). The deletion of sugR resulted in increased mRNA levels of eno, pyk, and ldhA in acetate medium. Enzyme activity measurements revealed that SugR-mediated repression affects the activities of PfkA, Fba, and LdhA in vivo. As the deletion of sugR led to increased LdhA activity under aerobic and under oxygen deprivation conditions, L-lactate production by C. glutamicum was determined. The overexpression of sugR reduced L-lactate production by about 25%, and sugR deletion increased L-lactate formation under oxygen deprivation conditions by threefold. Thus, SugR functions as a global repressor of genes of the PTS, glycolysis, and fermentative L-lactate dehydrogenase in C. glutamicum.Corynebacterium glutamicum, which was isolated as an Lglutamate-excreting soil bacterium (1, 39), is a predominantly aerobic, biotin-auxotrophic, gram-positive bacterium widely used for the industrial production of more than 2 million tons of amino acids per year, mainly L-glutamate and L-lysine (31,43). A general view of this nonpathogenic bacterium, which has become a model organism for the Corynebacterineae, a suborder of Actinomycetales that also comprises the genus Mycobacterium (54), can be found in two recent monographs (9, 17).C. glutamicum is able to grow on a variety of sugars, sugar alcohols, and organic acids as sole carbon and energy sources (64). As in many other gram-positive and gram-negative bacteria, the phosphoenolpyruvate-dependent phosphotransferase system (PTS) is the major sugar uptake system (15,37,45,47). The PTS-mediated glucose, fructose, and sucrose uptake in C. glutamicum operates by phosphoryl group transfer from phosphoenolpyruvate via EI (encoded by ptsI) and HPr (ptsH) to the sugar-specific permeases EII Glc , EII Fru , and EII Suc , respectively (ptsG, ptsF, and ptsS, respectively).

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“…table 1, www.karger.com/doi/10.1159/000324502). Sprehe et al [2007] investigated the carbon catabolite repression in B. subtilis 168 and constructed pWH338 to obtain a chromosomal deletion of ccpA by substituting ccpA with the kanamycin resistance gene aphA3 . Due to the high similarity found between MA3.3 and B. subtilis 168, the previous construction made by Sprehe et al [2007] was used to delete ccpA in Bacillus sp.…”

Section: Construction and Characterization Of The Ccpa Mutantmentioning

confidence: 99%

“…MA3.3 [Rygus and Hillen, 1992] with pWH338 [Sprehe et al, 2007] was performed. In pWH338, the ccpA gene of B. subtilis 168 was substituted by the kanamycin resistance gene aphA3 [Sprehe et al, 2007]. For that reason, pWH338 was used to promote ccpA deletion by homologous recombination in Bacillus sp.…”

Section: Construction and Characterization Of The Ccpa Mutantmentioning

confidence: 99%

Role of <i>CcpA</i> in Polyhydroxybutyrate Biosynthesis in a Newly Isolated <i>Bacillus</i> sp. MA3.3

et al. 2011

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The ccpA gene was inactivated in the polyhydroxybutyrate (PHB)-producing strain Bacillus sp. MA3.3 in order to reduce glucose catabolite repression over pentoses and develop improved bacterial strains for the production of PHB from lignocellulosic hydrolysates. Mutant Bacillus sp. MSL7 ΔCcpA are unable to grow on glucose and ammonia as sole carbon and nitrogen sources, respectively. Supplementation of glutamate as the nitrogen source or the substitution of the carbon source by xylose allowed the mutant to partially recover its growth performance. RT-PCR showed that CcpA stimulates the expression of the operon (gltAB),responsible for ammonia assimilation via glutamate in Bacillus sp. MA3.3. Moreover, it was demonstrated that the supplementation of xylose or glutamate was capable of stimulating gltAB operon expression independently of CcpA. In PHB production experiments in mineral media, it has been observed that the glucose catabolite repression over the pentoses was partially released in MSL7. Although the carbohydrate consumption is faster in the ccpA mutant, the biomass and PHB biosynthesis are lower, even with supplementation of glutamate. This is attributed to an increase of acetyl-CoA flux towards the tricarboxylic acid cycle observed in the mutant.

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CcpA Mutants with Differential Activities in <i>Bacillus subtilis</i>

Cited by 18 publications

References 61 publications

Distinct Time-Resolved Roles for Two Catabolite-Sensing Pathways during Streptococcus pyogenes Infection

Distinct Time-Resolved Roles for Two Catabolite-Sensing Pathways during Streptococcus pyogenes Infection

The Global Repressor SugR Controls Expression of Genes of Glycolysis and of the l -Lactate Dehydrogenase LdhA in Corynebacterium glutamicum

Role of <i>CcpA</i> in Polyhydroxybutyrate Biosynthesis in a Newly Isolated <i>Bacillus</i> sp. MA3.3

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