1887

Abstract

Ammonia assimilation in biovar strain RCR1001 (hereafter called ) appears to take place only through the glutamine synthetase/glutamate synthase pathway since () no glutamate dehydrogenase was detected in crude extracts of bacteria grown in different nitrogen sources, and () the growth rate on glutamine as a nitrogen source was faster than that observed on NHCl. In contrast to reports for other species, can definitely utilize NHC1 for growth. contains two glutamine synthetase isoenzymes, GSI and GSII, which can be detected in the presence of each other by differential heat stability, or separated by affinity chromatography or immunoabsorption with an antiserum raised against pure GSI. GSII does not cross-react with an anti-GSI antiserum. GSI was shown to be reversibly adenylylated and it was also shown that adenylylation inhibits the biosynthetic activity of this enzyme, in a similar way to that reported for glutamine synthetase and in contrast to that observed for glutamine synthetase of sp. strain ANU289. The apparent adenylylation level in different growth conditions changes from 21% to 99%, indicating a physiological role of this post-translational modification in the regulation of GSI activity. The intracellular concentration of GSI varies very little when is grown on different nitrogen sources (twofold when measured by the transferase assay, or fourfold when measured by ELISA). In addition, the concentration of mRNA specific for GSI in different nitrogen sources does not show appreciable differences. The intracellular concentration of GSII varies from a specific activity value higher than 1000 when is grown on glutamate or nitrate, to an undetectabie level when grown on NHC1. When NHC1 is added to a culture growing in glutamate, GSII activity is rapidly diluted out, suggesting a post-translational mechanism of enzyme inhibition or inactivation. Chloramphenicol prevents the disappearance of GSII activity, thus suggesting that protein synthesis is required for this process.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-3-629
1989-03-01
2024-12-09
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/3/mic-135-3-629.html?itemId=/content/journal/micro/10.1099/00221287-135-3-629&mimeType=html&fmt=ahah

References

  1. Bender R.A., Janssen K.A., Resnick A.D., Blumenberg M., Foor F., Magasanik B. 1977; Biochemical parameters of glutamine synthetase from Klebsiella aerogenes . Journal of Bacteriology 129:1001–1009
    [Google Scholar]
  2. Bravo A., Mora J. 1988; Ammonium assimilation in Rhizobium phaseoli by the glutamine synthetase-glutamate synthase pathway. Journal of Bacteriology 170:980–984
    [Google Scholar]
  3. Carlson T.A., Chelm B.K. 1986; Apparent eukaryotic origin of GSII from the bacterium B. japonicum . Nature; London: 322568–570
    [Google Scholar]
  4. Carlson T.A., Guerinot M., Chelm B.K. 1985; Characterization of the gene encoding GSI (glnA) from B. japonicum . Journal of Bacteriology 162:698–703
    [Google Scholar]
  5. Carlson T.A., Martin G.B., Chelm B.K. 1987; Differential transcription of the two glutamine synthetase genes of B. japonicum . Journal of Bacteriology 169:5861–5865
    [Google Scholar]
  6. Colonna-Romano S., Riccio A., Guida M., Defez R., Lamberti A., Iaccarino M., Arnold W., Priefer U., PÜhler A. 1987; Tight linkage of glnA and a putative regulatory gene in R. leguminosarum . Nucleic Acid Research 15:1951–1963
    [Google Scholar]
  7. Darrow R. 1980; Role of glutamine synthetase in nitrogen fixation. In Glutamine Synthetase: Metabolism, Enzymology and Regulation pp. 139–166 Mora J., Palacios R. Edited by New York: Academic Press;
    [Google Scholar]
  8. Darrow R.A., Knotts R.R. 1977; Two forms of glutamine synthetase in free-living root-nodule bacteria. Biochemical and Biophysical Research Communications 78:554–559
    [Google Scholar]
  9. Dixon R. 1984; Tandem promoters determine regulation of the K. pneumoniae glutamine synthetase(glnA) gene. Nucleic Acids Research 12:7811–7830
    [Google Scholar]
  10. Engvall E. 1980; Enzyme immunoassay ELISA and EMIT. Methods in Enzymology 70:419–439
    [Google Scholar]
  11. Ferguson A.R., Sims A.P. 1971; Inactivation in vivo of glutamine synthetase and NAD-specific glutamate dehydrogenase: its role in regulation of glutamine synthesis in yeast. Journal of General Microbiology 69:423–427
    [Google Scholar]
  12. Filser M.M.K., Moscatelli C., Lamberti A., Vincze E., Guida M., Salzano G., Iaccarino M. 1986; Characterization and cloning of two Rhizobium leguminosarum genes coding for glutamine synthetase activities. Journal of General Microbiology 132:2561–2569
    [Google Scholar]
  13. Fuchs R.L., Keister D.L. 1980; Comparative properties of glutamine synthetases I and II in Rhizobium and Agrobacterium spp. Journal of Bacteriology 144:641–648
    [Google Scholar]
  14. Gussin G.N., Ronson C.W., Ausubel F.M. 1986; Regulation of nitrogen fixation genes. Annual Review of Genetics 20:567–591
    [Google Scholar]
  15. Hooykaas P.J.J., Klapwijk P.M., Nuti M.P., Schilperoort R.A., Roersch A. 1977; Transfer of the Agrobacterium tumefaciens TI plasmid to avirulent agrobacteria and to Rhizobium ex planta. Journal of General Microbiology 98:477–484
    [Google Scholar]
  16. Howitt S.M., Gresshoff P.M. 1985; Ammonia regulation of glutamine synthetase in Rhizobium sp. ANU289. Journal of General Microbiology 131:1433–1440
    [Google Scholar]
  17. Ludwig R.A. 1978; Control of ammonium assimilation in Rhizobium 32H1. Journal of Bacteriology 135:114–123
    [Google Scholar]
  18. Ludwig R.A. 1980; Physiological roles of glutamine synthetases I and II in ammonium assimilation in Rhizobium sp. 32H1. Journal of Bacteriology 141:1209–1216
    [Google Scholar]
  19. Melton D.A., Krieg P.A., Rebagliati M.R., Maniatis T., Zinn K., Green M.R. 1984; Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Research 12:7035–7056
    [Google Scholar]
  20. Morett E., Moreno S., Espin G. 1985; Impaired nitrogen fixation and glutamine synthesis in methionine sulfoximine sensitive (MSs) mutants of Rhizobium phaseoli . Molecular and General Genetics 200:229–234
    [Google Scholar]
  21. Prusiner S., Miller R.E., Valentine R.C. 1972; Adenosine 3′5′-cyclic monophosphate control of the enzymes of glutamine metabolism in E. coli . Proceedings of the National Academy of Sciences of the United States of America 69:2922–2926
    [Google Scholar]
  22. Shapiro B.M., Stadtman E.R. 1970; The regulation of glutamine synthetase in microorganisms. Annual Review of Microbiology 24:501–524
    [Google Scholar]
  23. Somerville E., Kahn M.L. 1983; Cloning of the GSI gene from R. meliloti . Journal of Bacteriology 156:168–176
    [Google Scholar]
  24. Stadtman E.R., Ginsburg A. 1974; The glutamine synthetase of E. coli: structure and control. In The Enzymes X pp. 755–807 Boyer P. D. Edited by New York: Academic Press;
    [Google Scholar]
  25. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4354
    [Google Scholar]
/content/journal/micro/10.1099/00221287-135-3-629
Loading
/content/journal/micro/10.1099/00221287-135-3-629
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error