Reference : Overexpression of a Streptomyces viridochromogenes gene (glnII) encoding a glutamine ...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
Overexpression of a Streptomyces viridochromogenes gene (glnII) encoding a glutamine synthetase similar to those of eucaryotes confers resistance against the antibiotic phosphinothricyl-alanyl-alanine
Behrmann, Iris mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit >]
Hillemann, D. [> >]
Pühler, A. [> >]
Strauch, E. [> >]
Wohlleben, W. [> >]
Journal of Bacteriology
American Society for Microbiology (ASM)
Yes (verified by ORBilu)
[en] Amino Acid Sequence ; Sequence Homology, Nucleic Acid ; Restriction Mapping ; Replicon ; Plasmids ; Organophosphorus Compounds ; Molecular Sequence Data ; Glutamate-Ammonia Ligase ; Genes, Bacterial ; Gene Expression ; Drug Resistance, Microbial ; DNA, Bacterial ; Blotting, Southern ; Base Sequence ; Anti-Bacterial Agents ; Streptomyces
[en] Phosphinothricyl-alanyl-alanine (PTT), also known as bialaphos, contains phosphinothricin, a potent inhibitor of glutamine synthetase (GS). A 2.75-kilobase NcoI fragment of the Streptomyces viridochromogenes PTT-resistant mutant ES2 cloned on a multicopy vector mediated PTT resistance to S. lividans and to S. viridochromogenes. Nucleotide sequence analysis of the 2.75-kb NcoI fragment revealed the presence of three open reading frames. Open reading frame 3 was termed glnII since significant similarity was found between its deduced amino acid sequence and those from GS of eucaryotes and GSII of members of the family Rhizobiaceae. Subcloning experiments showed that PTT resistance is mediated by overexpression of glnII encoding a 37.3-kilodalton protein of 343 amino acids. A three- to fourfold increase in gamma-glutamyltransferase activity could be observed in S. lividans transformants carrying the glnII gene on a multicopy plasmid. For S. viridochromogenes it was shown that PTT resistance conferred by the 2.75-kb NcoI fragment was dependent on its multicopy state. GS activity encoded by glnII was found to be heat labile. Southern hybridization with seven different Streptomyces strains suggested that they all carry two types of GS genes, glnA and glnII.

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