1887

Abstract

SUMMARY: The structural gene () encoding the glutamine synthetase (GS) of the extremely thermophilic eubacterium has been cloned on a 6·0 kb III DNA fragment. Sequencing of the region containing the gene (1444 bp) showed an ORF encoding a polypeptide (439 residues) with an estimated mass of 50088 Da, which shared significant homology with the GSI sequences of other Bacteria () and Archaea (). The gene was expressed in , as shown by the ability to complement a lesion in the glutamine-auxotrophic strain ET8051. The recombinant GS has been partially characterized with respect to the temperature dependence of enzyme activity, molecular mass and mode of regulation. The molecular mass of the GS (590000 Da), estimated by gel filtration, was compatible with a dodecameric composition for the holoenzyme, as expected for a glutamine synthetase of the GSI type. Comparison of the amino acid sequence of GS with those from thermophilic and mesophilic micro-organisms failed to detect any obvious features directly related to thermal stability.

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1992-02-01
2021-10-28
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References

  1. Almassy R. J., Janson C. A., Hamlin R., Xuong N.-H., Eisenberg D. 1986; Novel subunit-subunit interactions in the structure of glutamine synthetase. Nature. London 323:304–309
    [Google Scholar]
  2. Behrmann I., Hlllemann D., Puhler A., Strauch E., Wohlleben W. 1990; Overexpression of a Streptomyces virido-chromogenes gene (gMI) encoding a glutamine synthetase similar to those of eukaryotes confers resistance against the antibiotic phosphinothricyl-alanyl-alanine. Journal of Bacteriology 172:5326–5334
    [Google Scholar]
  3. Bhatnagar L., Zeikus J. G., Aubert J.-P. 1986; Purification and characterization of glutamine synthetase from the archaebacterium Methanobacterium ivanovi. Journal of Bacteriology 165:638–643
    [Google Scholar]
  4. Blin N., Stafford D. W. 1976; A general method for the isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Research 3:2303–2308
    [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C, Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. 1977; Construction and characterization of new cloning vehicles II. A multipurpose cloning system. Gene 2:95–113
    [Google Scholar]
  6. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  7. Branlant C, Oster T., Branlant G. 1989; Nucleotide sequence determination of the DNA region coding for Bacillus stearothermo-philus glyceraldehyde-3-phosphate dehydrogenase and of the flanking DNA regions required for its expression in Escherichia coli. Gene 75:145–155
    [Google Scholar]
  8. Carlson T. A., Chelm B. K. 1986; Apparent eukaryotic origin of glutamine synthetase II from the bacterium Bradyrhizobium japonicum. Nature, London 322:568–570
    [Google Scholar]
  9. Colombo G., Villafranca J. J. 1986; Amino acid sequence of Escherichia coli glutamine synthetase deduced from the DNA nucleotide sequence. Journal of Biological Chemistry 261:10587–10591
    [Google Scholar]
  10. Cubellis M. V., Rozzo C, Montecucchi P., Rossi M. 1990; Isolation and sequencing of a new /f-galactosidase-encoding archae-bacterial gene. Gene 94:89–94
    [Google Scholar]
  11. Cullimore J. V., Bennet M. J. 1988; The molecular biology and biochemistry of plant glutamine synthetase from root nodules of Phaseolus vulgaris L. and other legumes. Journal of Plant Physiology 132:387–393
    [Google Scholar]
  12. Dagert M., Ehrlich S. D. 1979; Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28
    [Google Scholar]
  13. Deuel T. F., Prusiner S. 1974; Regulation of glutamine synthetase from Bacillus subtilis by divalent cations, feedback inhibitors, and L-glutamine. Journal of Biological Chemistry 249:257–264
    [Google Scholar]
  14. Fabry S., Lang J., Niermann T., Vingron M., Hensel R. 1989; Nucleotide sequence of the glyceraldehyde-3-phosphate dehydrogenase gene from the mesophilic methanogenic archae-bacteria Methanobacterium bryantii and Methanobacterium formici-cum. Comparison with the respective gene structure of the closely related extreme thermophile Methanothermus fervidus. European Journal of Biochemistry 179:405–413
    [Google Scholar]
  15. Fisher R., Tuli R., Haselkorn R. 1981 A cloned cyanobacterial gene for glutamine synthetase functions in Escherichia coli, but the enzyme is not adenylylated. Proceedings of the National Academy of Sciences of the United States of America 783393–3397
  16. Fisher S. H., Wray L. V. 1989; Regulation of glutamine synthetase in Streptomyces coelicolor. Journal of Bacteriology 171:2378–2383
    [Google Scholar]
  17. Haizer D. J., Moses V. 1978; The enzymes of proline biosynthesis in Escherichia coli. Their molecular weights and the problem of enzyme aggregation. Biochemical Journal 173:219–228
    [Google Scholar]
  18. Harris J. I., Perham R. N. 1968; Glyceraldehyde 3-phosphate dehydrogenase from pig muscle. Nature, London 219:1025–1028
    [Google Scholar]
  19. Higuchi R., Stang H. D., Browne J. K., Martin M. O., Huot M., Lipeles J., Salser W. 1981; Human ribosomal RNA gene spacer sequences are found interspersed elsewhere in the genome. Gene 15:177–186
    [Google Scholar]
  20. Hill R. T., Parker J. R., Goodman H. J. K., Jones D. T., Woods D. R. 1989; Molecular analysis of a novel glutamine synthetase of the anaerobe Bacteroides fragilis. Journal of General Microbiology 135:3271–3279
    [Google Scholar]
  21. Huber R., Langworthy T. A., Konig H., Thomm M., Woese C. R., Sleytr U. B., Stetter K. O. 1986; Thermotoga maritima sp. nov. represents a new genus of unique extremely thermophilic eubacteria growing up to 90 °C. Archives of Microbiology 144:324–333
    [Google Scholar]
  22. Janssen P. J., Jones W. A., Jones D. T., Woods D. R. 1988; Molecular analysis and regulation of the glnA gene of the Gram-positive anaerobe Clostridium acetobutylicum. Journal of Bacteriology 170:400–408
    [Google Scholar]
  23. Kumada Y., Takano E., Nagaoka K., Thompson C. J. 1990; Streptomyces hygroscopicus has two glutamine synthetase genes. Journal of Bacteriology 172:5343–5351
    [Google Scholar]
  24. Love D. R., Streiff M. B. 1987; Molecular cloning of a β-glucosidase gene from an extremely thermophilic anaerobe in E. coli and B. subtilis. Biotechnology 5:384–387
    [Google Scholar]
  25. Love D. R., Fisher R., Bergquist P. L. 1988; Sequence structure and expression of a cloned β-glucosidase gene from an extreme thermophile. Molecular General Genetics 213:84–92
    [Google Scholar]
  26. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  27. Marck C. 1988; 'DNA Strider': a 'C program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Research 16:1829–1836
    [Google Scholar]
  28. Nakano Y., Tanaka E., Kato C, Kimura K., Horikoshi K. 1989; The complete nucleotide sequence of the glutamine synthetase gene (glnA) of Bacillus subtilis. FEMS Microbiology Letters 57:81–86
    [Google Scholar]
  29. Pledger R. J., Baross J. A. 1991; Preliminary description and nutritional characterization of a chemoorganotrophic archaeo-bacterium growing at temperatures of up to 110 °C isolated from a submarine hydrothermal vent environment. Journal of General Microbiology 137:203–211
    [Google Scholar]
  30. Possot O., Sibold L., Aubert J.-P. 1989; Nucleotide sequence and expression of the glutamine synthetase structural gene, glnA, of the archaebacterium Methanococcus voltae. Research in Microbiology 140:355–371
    [Google Scholar]
  31. Rawlings D. E., Jones W. A., O'Neill E. G., Woods D. R. 1987; Nucleotide sequence of the glutamine synthetase gene and its controlling region from the acidophilic autotroph Thiobacillus ferrooxidans. Gene 53:211–217
    [Google Scholar]
  32. Reitzer L. J., Magasanik B. 1987 Ammonia assimilation and the biosynthesis of glutamine, glutamate, aspartate, asparagine, L-alanine and d-alanine. In Escherichia coli and Salmonella typhimurium, pp. 302–320 Edited by Neidhart F. C. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  33. Riccardi G., De Rossi E., Della Valle G., Ciferri O. 1985; Cloning of the glutamine synthetase gene from Spirulina platensis. Plant Molecular Biology 4:133–136
    [Google Scholar]
  34. Sanangelantoni A. M., Barbarini D., Di Pasquale G., Cammarano P., Tiboni O. 1990; Cloning and nucleotide sequence of an archaebacterial glutamine synthetase gene: phylo-genetic implications. Molecular General Genetics 221:187–194
    [Google Scholar]
  35. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 745463–5467
  36. Schultes V., Deutzmann R., Jaenicke R. 1990; Complete amino-acid sequence of glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic eubacterium Thermotoga maritima. European Journal of Biochemistry 192:25–31
    [Google Scholar]
  37. Shapiro B. M., Stadtman E. R. 1968; 5′-Adenylyl-0-tyrosine. The novel phosphodiester residue of adenylylated glutamine synthetase from Escherichia coli. Journal of Biological Chemistry 243:3769–3771
    [Google Scholar]
  38. Shapiro B. M., Kingdom H. S., Stadtman E. R. 1967; Regulation of glutamine synthetase, VII. Adenylyl glutamine synthetase: a new form of the enzyme with altered regulatory and kinetic properties. Proceedings of the National Academy of Sciences of the United States of America 58642–649
  39. Shatters R. G., Kahn M. L. 1989; Glutamine synthetase II in Rhizobium: reexamination of the proposed horizontal transfer of DNA from eukaryotes to prokaryotes. Journal of Molecular Evolution 29:422–428
    [Google Scholar]
  40. Shine C., Dalgarno L. 1976; Determinant of cistron specificity in bacterial ribosomes. Nature, London 254:34–38
    [Google Scholar]
  41. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  42. Stadtman E. R., Smyrniotis P. Z., Davis J. N., Wittemberg M. E. 1979; Enzymic procedures for determining the average state of adenylylation of Escherichia coli glutamine synthetase. Analytical Biochemistry 95:275–285
    [Google Scholar]
  43. Streicher S. L., Tyler B. 1980; Purification of glutamine synthetase from a variety of bacteria. Journal of Bacteriology 142:69–78
    [Google Scholar]
  44. Streicher S. L., Tyler B. 1981; Regulation of glutamine synthetase activity by adenylylation in the Gram-positive bacterium Streptomyces cattleya. Proceedings of the National Academy of Sciences of the United States of America 78229–233
  45. Tiboni O., Sanangelantoni A. M., Cammarano P., Cimino L., Di Pasquale G., Sora S. 1989; Expression in Escherichia coli ofthe tuf gene from the extremely thermophilic eubacterium Thermotoga maritima : Purification of the Thermotoga elongation factor Tu by thermal denaturation of the mesophile host cell proteins. Systematic and Applied Microbiology 12:127–133
    [Google Scholar]
  46. Toukdarian A., Saunders G., Selman-Sosa G., Santero E., Woodley P., Kennedy C. 1990; Molecular analysis of the Azotobacter vinelandii glnA gene encoding glutamine synthetase. Journal of Bacteriology 172:6529–6539
    [Google Scholar]
  47. Tyler B. 1978; Regulation of the assimilation of nitrogen compounds. Annual Review of Biochemistry 47:1127–1162
    [Google Scholar]
  48. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
    [Google Scholar]
  49. Wilbur W. J., Lipman D. J. 1983; Rapid similarity searches of nucleic acid and protein data banks. Proceedings of the National Academy of Sciences of the United States of America 80726–730
  50. Woese C. R. 1987; Bacterial evolution. Microbiological Reviews 51:221–271
    [Google Scholar]
  51. Wray L. V., Fisher S. H. 1988; Cloning and nucleotide sequence of the Streptomyces coelicolor gene encoding glutamine synthetase. Gene 71:247–256
    [Google Scholar]
  52. Zillig W., Holz I., Janekovic D., Klenk H.-D., Imsel E., Trent J., Wunderl S., Forjaz V. H., CoUTINHO R., Ferreira T. 1990; Hyperthermias butylicus, a hyperthermophilic sulfur-reducing archaebacterium that ferments peptides. Journal of Bacteriology 172:3959–3965
    [Google Scholar]
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