An<i>Escherichia coli</i>protein with homology to the H-protein of the glycine cleavage enzyme complex from pea and chicken liver

Stauffer, Lorraine T.; Steiert, Paula S.; Steiert, John G.; Stauffer, George V.

doi:10.3109/10425179109008434

Cited by 9 publications

(4 citation statements)

References 22 publications

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“…2). The nucleotide sequence is identical to that reported for an E. coli gene encoding H-protein [26]. The molecular mass of the E. coli H-protein was calculated to be 13 679 Da which is significantly lower than the value (17.3 kDa) estimated by SDS/PAGE (Fig.…”

Section: Dna Sequence Analysissupporting

confidence: 63%

“…The system is composed of four proteins named p-, H-, T-and L-protein In Escherichia coli the glycine-cleavage system is inducible by exogenous glycine [24] and encoded by the gcv structural genes situated at 62 min in the linkage map [25]. Although a gene for H-protein has been sequenced [26], the detailed properties of the glycine-cleavage system in E. coli have not been examined so far. Our purpose is to characterize the structural genes for the components of the glycine-cleavage system from E. coli and to acquire more information about the characteristic features of the deduced amino acid sequences and the binding sites for coenzymes and prosthetic groups.…”

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confidence: 99%

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Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine‐cleavage system

Okamura‐Ikeda

Ohmura

Fujiwara

et al. 1993

European Journal of Biochemistry

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) -EJB 930930/1 P-protein, H-protein and T-protein of the glycine cleavage system have been purified from Escherichia coli. Their N-terminal amino acid sequences were determined, and a set of oligonucleotide probes was designed for gene cloning. The nucleotide sequence of a fragment of DNA around the 62-min region of the E. coli chromosome, containing genes for the components of the glycinecleavage system has been determined. The sequence i?cludes three structural genes encoding Tprotein (363 amino acids, 40013 Da), H-protein (128 amino acids, 13679 Da) and P-protein (956 amino acids, 104240Da). These genes are named gcvT, gcvH and gcvP, respectively. They are organized in the above-mentioned order on the same strand of DNA with short intercistronic sequences. The presence of a potential promoter preceding gcvT and a typical rho-independent terminator sequence following gcvP indicated that the three genes constitute a single operon. Each component of the E. coli glycine-cleavage system exhibits considerable amino acid sequence similarity with the animal and plant counterparts. When the plasmid containing the gcv operon was transfected in E. coli cells, the gene products of gcvT gcvH and gcvP were overexpressed under the direction of the promoter of the gcv operon. However, bacteria harboring the plasmid that contained the gcv operon without the promoter region and the 5' terminal portion of gcvT failed to overexpress any of the three components.The glycine-cleavage system is a niultienzyme complex that catalyzes the reversible oxidation of glycine, yielding carbon dioxide, ammonia, methylenetetrahydrofolate and a reduced pyridine nucleotide [l -51. The enzyme system has been purified from a number of sources including animal liver [6], plants [7,8] and bacteria such as Pepfococcus glycinophilus [9], Arthrobacter globformis [lo] and Eubacterium acidaminophilum [ll]. The system is composed of four proteins named p-, H-, T-and L-protein In Escherichia coli the glycine-cleavage system is inducible by exogenous glycine [24] and encoded by the gcv structural genes situated at 62 min in the linkage map [25]. Although a gene for H-protein has been sequenced [26], the detailed properties of the glycine-cleavage system in E. coli have not been examined so far. Our purpose is to characterize the structural genes for the components of the glycine-cleavage system from E. coli and to acquire more information about the characteristic features of the deduced amino acid sequences and the binding sites for coenzymes and prosthetic groups. In this study, we report the molecular cloning of these genes, their nucleotide sequences and deduced amino acid sequences and evidence for their arrangement in an operon. Where possible, these proteins have been compared to their animal and plant counterparts.

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Section: Dna Sequence Analysissupporting

confidence: 63%

mentioning

confidence: 99%

Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine‐cleavage system

Okamura‐Ikeda

Ohmura

Fujiwara

et al. 1993

European Journal of Biochemistry

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“…1) . This fragment i s believed tlo encode the entire E. coli GCV system, except the L-protein (Stauffer et a/., 1986;Steiert et a/., 1991).…”

Section: Resultsmentioning

confidence: 99%

TheEscherichia coli gcvTgene encoding the T-protein of the glycine cleavage enzyme system

1993

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Plasmid pGS146 carries the Escherichia coli gcv system on a 7.12 kb SalI-BamHI DNA insert fragment. The DNA sequence of a gene which presumably encodes the T-protein of the glycine cleavage (GCV) enzyme complex was determined. The gene, designated gcvT, encodes a polypeptide of 364 amino acids with a calculated molecular weight of 40,146 daltons. In a minicell system, the SalI-BamHI fragment directs the synthesis of three polypeptides with Mr values of about 93,300, 43,300 and 17,400 daltons. When gcvT was inactivated by insertion of a translation terminator sequence, the Mr 43,300 dalton polypeptide was not observed. The deduced amino acid sequence of the E. coli T-protein was compared with the sequence of the T-protein from bovine liver. 190 of 364 amino acid residues are identical or chemically similar between the two proteins. An S1 nuclease mapping experiment located the transcription start point for gcvT. Single basepair changes were made in the promoter -10 and -35 sequences. These mutations significantly reduced expression from a gcvT-lacZ gene fusion. The gcvT gene is transcribed and translated in the same direction as the gcvH gene.

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“…A number of genes involved in metabolism were also dysregulated by SK-03-92 treatment, including the gcvH gene that encodes GcvH, which shuttles the methylamine group of glycine from the P-protein to the T-protein via a lipoyl group [ 46 ]. Genes associated with protein degradation and repair had altered transcript abundance in SK-03-92-treated S. aureus .…”

Section: Resultsmentioning

confidence: 99%

Identification of Staphylococcus aureus Cellular Pathways Affected by the Stilbenoid Lead Drug SK-03-92 Using a Microarray

et al. 2017

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The mechanism of action for a new lead stilbene compound coded SK-03-92 with bactericidal activity against methicillin-resistant Staphylococcus aureus (MRSA) is unknown. To gain insight into the killing process, transcriptional profiling was performed on SK-03-92 treated vs. untreated S. aureus. Fourteen genes were upregulated and 38 genes downregulated by SK-03-92 treatment. Genes involved in sortase A production, protein metabolism, and transcriptional regulation were upregulated, whereas genes encoding transporters, purine synthesis proteins, and a putative two-component system (SACOL2360 (MW2284) and SACOL2361 (MW2285)) were downregulated by SK-03-92 treatment. Quantitative real-time polymerase chain reaction analyses validated upregulation of srtA and tdk as well as downregulation of the MW2284/MW2285 and purine biosynthesis genes in the drug-treated population. A quantitative real-time polymerase chain reaction analysis of MW2284 and MW2285 mutants compared to wild-type cells demonstrated that the srtA gene was upregulated by both putative two-component regulatory gene mutants compared to the wild-type strain. Using a transcription profiling technique, we have identified several cellular pathways regulated by SK-03-92 treatment, including a putative two-component system that may regulate srtA and other genes that could be tied to the SK-03-92 mechanism of action, biofilm formation, and drug persisters.

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AnEscherichia coliprotein with homology to the H-protein of the glycine cleavage enzyme complex from pea and chicken liver

Cited by 9 publications

References 22 publications

Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine‐cleavage system

Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine‐cleavage system

TheEscherichia coli gcvTgene encoding the T-protein of the glycine cleavage enzyme system

Identification of Staphylococcus aureus Cellular Pathways Affected by the Stilbenoid Lead Drug SK-03-92 Using a Microarray

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