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Your search keyword '"metabolism [Nucleotides]"' showing total 2 results
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2 results on '"metabolism [Nucleotides]"'

1. Nucleotide flips determine the specificity of the Ecl18kI restriction endonuclease

Author

Matthias Bochtler, Elena Manakova, Saulius Grazulis, Gintautas Tamulaitis, Honorata Czapinska, Roman H. Szczepanowski, and Virginijus Siksnys

Subjects
Models, Molecular, Base pair, Macromolecular Substances, Molecular Sequence Data, Cleavage (embryo), Crystallography, X-Ray, General Biochemistry, Genetics and Molecular Biology, Article, Substrate Specificity, Endonuclease, chemistry.chemical_compound, genetics [Deoxyribonucleases, Type II Site-Specific], ddc:570, Nucleotide, Amino Acid Sequence, chemistry [Deoxyribonucleases, Type II Site-Specific], Binding site, Deoxyribonucleases, Type II Site-Specific, Molecular Biology, Peptide sequence, chemistry.chemical_classification, chemistry [DNA], Binding Sites, metabolism [Deoxyribonucleases, Type II Site-Specific], General Immunology and Microbiology, biology, Base Sequence, Nucleotides, General Neuroscience, DNA, Protein Structure, Tertiary, metabolism [Nucleotides], Restriction enzyme, chemistry [Nucleotides], Biochemistry, chemistry, endodeoxyribonuclease Ecl18kI, biology.protein, metabolism [DNA], Nucleic Acid Conformation, Dimerization
Abstract

Restricion endonuclease Ecl18kI is specific for the sequence /CCNGG and cleaves it before the outer C to generate 5 nt 5′-overhangs. It has been suggested that Ecl18kI is evolutionarily related to NgoMIV, a 6-bp cutter that cleaves the sequence G/CCGGC and leaves 4 nt 5′-overhangs. Here, we report the crystal structure of the Ecl18kI–DNA complex at 1.7 A resolution and compare it with the known structure of the NgoMIV–DNA complex. We find that Ecl18kI flips both central nucleotides within the CCNGG sequence and buries the extruded bases in pockets within the protein. Nucleotide flipping disrupts Watson–Crick base pairing, induces a kink in the DNA and shifts the DNA register by 1 bp, making the distances between scissile phosphates in the Ecl18kI and NgoMIV cocrystal structures nearly identical. Therefore, the two enzymes can use a conserved DNA recognition module, yet recognize different sequences, and form superimposable dimers, yet generate different cleavage patterns. Hence, Ecl18kI is the first example of a restriction endonuclease that flips nucleotides to achieve specificity for its recognition site.

Published
2006

2. Phosphate starvation-inducible gene ushA encodes a 5 ' nucleotidase required for growth of Corynebacterium glutamicum on media with nucleotides as the phosphorus source

Author

Volker F. Wendisch, Doris Rittmann, and Ulrike Sorger-Herrmann

Subjects
ushA protein, E coli, promoters, Applied Microbiology and Biotechnology, 5'-nucleotidase, Corynebacterium glutamicum, chemistry.chemical_compound, Environmental Microbiology, metabolism [5'-Nucleotidase], Nucleotide, 5'-Nucleotidase, chemistry [Phosphoric Diester Hydrolases], chemistry.chemical_classification, Ecology, Nucleotides, enzymology [Corynebacterium glutamicum], Escherichia coli Proteins, Phosphorus, genetics [5'-Nucleotidase], metabolism [Phosphoric Diester Hydrolases], growth & development [Corynebacterium glutamicum], metabolism [Phosphorus], metabolism [Adenosine Monophosphate], Biochemistry, salmonella-enterica, genetics [Escherichia coli Proteins], Starvation response, genetics [Bacterial Proteins], Biotechnology, transcriptional analysis, plasmids, chemistry [Bacterial Proteins], metabolism [Bacterial Proteins], Molecular Sequence Data, cloning, Biology, metabolism [Escherichia coli Proteins], Phosphates, Phosphorus metabolism, Bacterial Proteins, ddc:570, Nucleotidase, expression, molecular analysis, metabolism [Phosphates], genetics [Phosphoric Diester Hydrolases], Sugar phosphates, Base Sequence, Phosphoric Diester Hydrolases, transformation, genetics [Corynebacterium glutamicum], Gene Expression Regulation, Bacterial, sequence, Uridine Diphosphate Sugars, Phosphate, Adenosine Monophosphate, metabolism [Nucleotides], chemistry, escherichia-coli, bacteria, chemistry [5'-Nucleotidase], metabolism [Uridine Diphosphate Sugars], Food Science
Abstract

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.

Published
2005

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