Promoters of Corynebacterium glutamicum

Pátek, Miroslav; Nešvera, Jan; Guyonvarch, Armel; Reyes, Oscar; Leblon, Gérard

doi:10.1016/s0168-1656(03)00155-x

Cited by 131 publications

(119 citation statements)

References 56 publications

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“…In four independently obtained pDrive ramBpRACE clones, the TS of ramB was found to be an A residue which is identical to the translational start site proposed (11) (NCBI accession number NC_006958). Such leaderless transcripts are not uncommon in C. glutamicum and other actinomycetes (16). Inspection of the upstream region of the TS led to the identification of a potential Ϫ10 box (TATAGT) which is conserved in five of six nucleotides relative to the Ϫ10 consensus sequence described for corynebacteria (TA[C/T]AAT) (15).…”

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confidence: 99%

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RamB, the Transcriptional Regulator of Acetate Metabolism in Corynebacterium glutamicum , Is Subject to Regulation by RamA and RamB

et al. 2007

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In Corynebacterium glutamicum, the transcriptional regulator RamB negatively controls the expression of genes involved in acetate metabolism. Here we show that RamB represses its own expression by direct interaction with a 13-bp motif in the ramB promoter region. Additionally, ramB expression is subject to carbon source-dependent positive control by RamA.Corynebacterium glutamicum is an aerobic gram-positive soil bacterium well known for its use in large-scale biotechnological production of L-glutamate and lysine (10-14). We are interested in acetate metabolism and its regulation in this organism (7), and we recently identified and characterized two regulatory proteins, designated regulator of acetate metabolism A and B (RamA and RamB, respectively) from C. glutamicum ATCC 13032 (4, 6). RamA binds to single or tandem stretches of A/C/TG 4-6 T/C or AC 4-5 A/G/T and thereby acts as an activator of the pta, ack, aceA, and aceB genes, encoding phosphotransacetylase, acetate kinase, isocitrate lyase, and malate synthase, all involved in acetate metabolism of C. glutamicum. RamA was also shown to activate transcription of the surface layer protein gene (cspB) in C. glutamicum ATCC 14067 (9). Furthermore, RamA was shown to bind to the promoter region of its own gene and to act as a repressor there (3). The RamB protein specifically binds to a highly conserved 13-bp motif (AA/GAACTTTGCAAA) and represents a repressor of the pta, ack, aceA, and aceB genes when C. glutamicum is grown on glucose as a carbon and energy source (6). Inspection of the ramB promoter region revealed the presence of putative RamA and RamB binding sites, and this observation prompted us to study expression of ramB in cells growing in media containing glucose and/or acetate and to test for a regulatory function of RamA and RamB in ramB expression.The bacterial strains and the plasmids, their relevant characteristics and sources, and the oligonucleotides used in this study are given in Table 1. The media used and the methods not outlined explicitly (DNA preparation, promoter binding assays with hexahistidyl-tagged RamA and RamB fusion proteins, enzyme assays, RNA preparation and identification of the transcriptional start site by the RACE [rapid amplification of cDNA ends] method, generation of polyclonal antibodies, and Western blotting) were described previously (3, 4, 6).Western blot experiments with RamB-specific antibodies and crude extracts from C. glutamicum wild-type (WT) cells grown in minimal medium containing glucose, glucose plus acetate, and acetate showed that RamB is present in cells grown on either carbon source (Fig. 1). In agreement with the results of DNA affinity chromatography experiments (4), the largest amounts of RamB were observed in the cells grown on glucose alone.To test for ramB promoter activities in C. glutamicum cells grown in minimal media with glucose and/or acetate, we constructed reporter gene fusions by cloning a 593-bp ramB promoter fragment (covering the region 521 bp upstream to 72 bp downstream of the ramB sta...

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confidence: 99%

“…Inspection of the upstream region of the TS led to the identification of a potential Ϫ10 box (TATAGT) which is conserved in five of six nucleotides relative to the Ϫ10 consensus sequence described for corynebacteria (TA[C/T]AAT) (15). No apparent Ϫ35 region (TTGCCA) could be recognized, which also is a common feature of C. glutamicum promoters (15,16).…”

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confidence: 99%

RamB, the Transcriptional Regulator of Acetate Metabolism in Corynebacterium glutamicum , Is Subject to Regulation by RamA and RamB

et al. 2007

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“…Some bacterial species have been found to possess promoters that have conserved hexanucleotide sequences that are different from the E. coli consensus sequences, and the promoter regions of some bacterial species do not appear to have conserved regions [36,37].…”

Section: Figure 21 Strategy For the Replacement Of The Carba Carbbmentioning

confidence: 99%

“…The desulfurization enzymes might be the rate-limiting factors in achieving desulfurization activity up to a certain level, but in order to achieve even higher activities alterations in genes and regulatory sequences other than (or in addition to) the dsz genes are needed. It is known from the study of the expression of metabolic pathways in other microbial species that alterations of transcriptional regulators/sequences as well as mutations in enzyme-encoding sequences are needed to enhance metabolic flow through a pathway [37]. Transcriptional regulators that are responsive to the concentration of various organosulfur compounds and that impact the expression of multiple genetic loci are known for some species [38], and an improved understanding of transcriptional and post-transcriptional gene regulation in Rhodococcus and other desulfurization-competent hosts will greatly benefit efforts to create biocatalysts with ever-higher desulfurization levels.…”

Section: Engineering Strains For Biodesulfurizationmentioning

confidence: 99%

Metabolic Engineering to Develop a Pathway for the Selective Cleavage of Carbon-Nitrogen Bonds

Kilbane¹

2006

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“…The promoter of ushA contains a conserved Ϫ10 region but lacks a conserved Ϫ35 region (Fig. 2); this was shown to be typical for C. glutamicum promoters (22,23).…”

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confidence: 96%

Phosphate Starvation-Inducible Gene ushA Encodes a 5′ Nucleotidase Required for Growth of Corynebacterium glutamicum on Media with Nucleotides as the Phosphorus Source

Rittmann

Sorger-Herrmann

Wendisch

2005

Appl Environ Microbiol

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Phosphorus is an essential component of macromolecules, like DNA, and central metabolic intermediates, such as sugar phosphates, and bacteria possess enzymes and control mechanisms that provide an optimal supply of phosphorus from the environment. UDP-sugar hydrolases and 5 nucleotidases may play roles in signal transduction, as they do in mammals, in nucleotide salvage, as demonstrated for UshA of Escherichia coli, or in phosphorus metabolism. The Corynebacterium glutamicum gene ushA was found to encode a secreted enzyme which is active as a 5 nucleotidase and a UDP-sugar hydrolase. This enzyme was synthesized and secreted into the medium when C. glutamicum was starved for inorganic phosphate. UshA was required for growth of C. glutamicum on AMP and UDP-glucose as sole sources of phosphorus. Thus, in contrast to UshA from E. coli, C. glutamicum UshA is an important component of the phosphate starvation response of this species and is necessary to access nucleotides and related compounds as sources of phosphorus.

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Promoters of Corynebacterium glutamicum

Cited by 131 publications

References 56 publications

RamB, the Transcriptional Regulator of Acetate Metabolism in Corynebacterium glutamicum , Is Subject to Regulation by RamA and RamB

RamB, the Transcriptional Regulator of Acetate Metabolism in Corynebacterium glutamicum , Is Subject to Regulation by RamA and RamB

Metabolic Engineering to Develop a Pathway for the Selective Cleavage of Carbon-Nitrogen Bonds

Phosphate Starvation-Inducible Gene ushA Encodes a 5′ Nucleotidase Required for Growth of Corynebacterium glutamicum on Media with Nucleotides as the Phosphorus Source

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