1887

Abstract

Summary: The deduced amino acid sequence derived from the sequence of a fragment of DNA from the free-living diazotroph was aligned to the homologous protein sequences encoded by the genes from and . High similarity was found in the central domain and in the C-terminal region. The putative NifA sequence was also found to contain an interdomain linker similar to that conserved among rhizobial NifA proteins, but not or . Analysis of the regulatory sequences found 5′ from indicated that the expression of this gene in is likely to be controlled by NifA, NtrC and RpoN, as judged by the presence of specific NifA- and NtrC-binding sites and characteristic −24/−12 promoters. Possible additional regulatory features included an ‘anaerobox’ and a site for integration host factor. The N-terminus of another open reading frame was found 3′ from and tentatively identified as by amino acid sequence comparison. The putative promoter sequence suggests that expression of may be activated by NifA and dependent on RpoN.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-7-1511
1991-07-01
2021-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/7/mic-137-7-1511.html?itemId=/content/journal/micro/10.1099/00221287-137-7-1511&mimeType=html&fmt=ahah

References

  1. Baldani, J. I.,, Baldani, V. L. D.,, Seldin, L., & Döbereiner, J.. ( 1986;). Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a root-associated nitrogen-fixing bacterium. . International Journal of Systematic Bacteriology 36, 86-93.
    [Google Scholar]
  2. Bankier, A. T.,, Weston, K. M., & Barrel, B. G.. ( 1987;). Random cloning and sequencing by the M13/dideoxynucleotide chain termination method. . Methods in Enzymology 155, 51-93.
    [Google Scholar]
  3. Bennett, L. T.,, Cannon, F., & Dean, D. R.. ( 1988;). Nucleotide sequence and mutagenesis of the nifA gene from Azotobacter vinelandii. . Molecular Microbiology 2, 315-321.
    [Google Scholar]
  4. Bozouklian, H., & Elmerich, C.. ( 1986;). Nucleotide sequence of the Azospirillum brasilense Sp7 glutamine synthetase structural gene. . Biochimie 68, 1181-1187.
    [Google Scholar]
  5. Bozouklian, H.,, Fogher, C., & Elmerich, C.. ( 1986;). Cloning and characterization of the glnA gene of Azospirillum brasilense Sp7. . Annales de l’Institut Pasteur 137B, 3-18.
    [Google Scholar]
  6. De Bruijn, F. J.,, Pawlowski, K.,, Ratet, P.,, Hilgert, U.,, Wong, C. H.,, Schneider, M.,, Meyer, H. Z. A., & Schell, J.. ( 1988;). Molecular genetics of nitrogen fixation by Azorhizobium caulinodans ORS571, the diazotrophic stem nodulating symbiont of Sesbania rostrata. . In Nitrogen Fixation: One Hundred Years After, pp. 351-355. Edited by H. Bothe,, F. J. de Bruijn, & W. E. Newton.. Stuttgart:: Gustav Fischer;.
    [Google Scholar]
  7. Buchanan-Wollaston, V.,., Cannon, M. C.,, Beynon, J. L., & Cannon, F. C.. ( 1981;). Role of the nifA gene product in the regulation of nif expression in Klebsiella pneumoniae. . Nature, London 294, 776-778.
    [Google Scholar]
  8. Buikema, W. J.,, Szeto, W. W.,, Lemley, P. V.,, Orme-Johnson, W. H., & Ausubel, F. M.. ( 1985;). Nitrogen fixation specific regulatory genes of Klebsiella pneumoniae and Rhizobium meliloti share homology with the general nitrogen regulatory gene ntrC of K. pneumoniae. . Nucleic Acids Research 13, 4539-4555.
    [Google Scholar]
  9. Buikema, W. J.,, Klingensmith, J. A.,, Gibbons, S. L., & Ausubel, F. M.. ( 1987;). Conservation of structure and location of Rhizobium meliloti and Klebsiella pneumoniae nifB genes. . Journal of Bacteriology 169, 1120-1126.
    [Google Scholar]
  10. Cannon, W. V.,, Kreutzer, R.,, Kent, H. M.,, Morett, E., & Buck, M.. ( 1990;). Activation of the Klebsiella nifU promoter: identification of multiple and overlapping upstream NifA binding sites. . Nucleic Acids Research 18, 1693-1701.
    [Google Scholar]
  11. Devereux, J.,, Haerberli, P., & Smithies, O.. ( 1984;). A comprehensive set of sequence analysis programs for the VAX. . Nucleic Acids Research 12, 387-395.
    [Google Scholar]
  12. Dixon, R.. ( 1988;). Genetic regulation of nitrogen fixation. . Symposia of the Society for General Microbiology 42, 417-438.
    [Google Scholar]
  13. Drummond, M.,, Clements, J.,, Merrick, M., & Dixon, R. A.. ( 1983;). Positive control and autogenous regulation of the nif LA promoter in Klebsiella pneumoniae. . Nature, London 301, 302-307.
    [Google Scholar]
  14. Drummond, M.,, Whitty, P., & Wootton, J.. ( 1986;). Sequence and domains relationship of ntrC and nifA from Klebsiella pneumoniae: homologies to other regulatory proteins. . EM BO Journal 5, 441-447.
    [Google Scholar]
  15. Falk, E. C.,, Johnson, J. L.,, Baldani, V. L. D.,, Döbereiner, J., & Krieg, N. R.. ( 1986;). Deoxyribonucleic and ribonucleic acid homology studies of the genera Azospirillum and Conglomeromonas. . International Journal of Systematic Bacteriology 36, 80-85.
    [Google Scholar]
  16. Feinberg, A. P., & Vogelstein, B.. ( 1984;). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. . Analytical Biochemistry 137, 266-267.
    [Google Scholar]
  17. Fischer, H. M., & Hennecke, H.. ( 1987;). Direct response of Bradyrhizobium japonicum nif A-mediated nif gene regulation to cellular oxygen status. . Molecular and General Genetics 209, 621-626.
    [Google Scholar]
  18. Fischer, H. M.,, Bruderer, T., & Hennecke, H.. ( 1988;). Essential and non essential domains in the Bradyrhizobium japonicum NifA protein : identification of indispensable cysteine residues potentially involved in redox reactivity and/or metal binding. . Nucleic Acids Research 16, 2207-2224.
    [Google Scholar]
  19. Goodrich, J. A.,, Schwartz, M. L., & McClure, W.. ( 1990;). Searching for and predicting the activity of sites for DNA binding proteins: compilation and analysis of the binding sites for Escherichia coli integration host factor (IHF). . Nucleic Acids Research 18,4993-5000.
    [Google Scholar]
  20. Grönger, P.,, Manian S. S.,, Reiländer, H.,, O’Connell, M.,, Priefer, U. B., & Pühler, A.. ( 1987;). Organization and partial sequence of a DNA region of the Rhizobium leguminosarum symbiotic plasmid pRL6JI containing the genes fixABC, nifA, nifB and a novel open reading frame. . Nucleic Acids Research 15, 31-49.
    [Google Scholar]
  21. Gussin, G. N.,, Ronson, C. W., & Ausubel, F. M.. ( 1986;). Regulation of nitrogen fixation genes. . Annual Review of Genetics 20, 567-591.
    [Google Scholar]
  22. Higgins, D. G., & Sharp, P. M.. ( 1988;). clustal: a package for performing multiple sequence alignment on a microcomputer. . Gene 73, 237-244.
    [Google Scholar]
  23. Hoover, T. R.,, Santero, E.,, Porter, S., & Kustu, S.. ( 1990;). The integration host factor stimulates interaction of RNA polymerase with NifA, the transcriptional activator for nitrogen fixation operons. . Cell 63, 11-22.
    [Google Scholar]
  24. Joerger, R. D., & Bishop, P. E.. ( 1988;). Nucleotide sequence and genetic analysis of the nifB-nifQ region from Azotobacter vinelandii. . Journal of Bacteriology 170, 1475-1487.
    [Google Scholar]
  25. Maniatis, T.,, Fritsch, E. F., & Sambrook, J.. ( 1982). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;.
    [Google Scholar]
  26. Masepohl, B.,, Klipp, W., & Pühler, A.. ( 1988;). Genetic characterization and sequence analysis of the duplicated nifA/nifB gene region of Rhodobacter capsulatus. . Molecular and General Genetics 212, 27-37.
    [Google Scholar]
  27. Mead, D. A.,, Szczena-Skorupa, E., & Kemper, B.. ( 1986;). Single stranded DNA ‘blue’ T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. . Protein Engineering 1, 67-74.
    [Google Scholar]
  28. Merrick, M. J.. ( 1983;). Nitrogen control of the nif regulon in Klebsiella pneumoniae: involvement of the ntrA gene and analogies between ntrC and nifA. . EM BO Journal 2, 39-44.
    [Google Scholar]
  29. Nees, D. W.,, Stein, P. A., & Ludwig, R. A.. ( 1988;). The Azorhizobium caulinodans nifA gene: identification of upstream activating sequences including a new element, the ‘anaerobox’. . Nucleic Acids Research 16, 9839-9853.
    [Google Scholar]
  30. Noti, J. D.,, Folkerts, O.,, Turken, A. N., & Szalay, A. A.. ( 1986;). Organization and characterization of genes essential for symbiotic nitrogen fixation from Bradyrhizobium japonicum I110. . Journal of Bacteriology 167, 774-783.
    [Google Scholar]
  31. Pedrosa, F. O., & Yates, M. G.. ( 1984;). Regulation of nitrogen fixation (nif) genes of Azospirillum brasilense by nifA and ntr (gin) type gene products. . FEMS Microbiology Letters 23, 95-101.
    [Google Scholar]
  32. Ronson, C. W.,, Nixon, B. T.,, Albright, C. M., & Ausubel, F. M.. ( 1987;). Rhizobium meliloti ntr A (rpoN) gene is required for diverse metabolic functions. . Journal of Bacteriology 169, 2424-2431.
    [Google Scholar]
  33. Santero, E.,, Toukdarian, A.,, Humphrey, R., & Kennedy, C.. ( 1988;). Identification and characterization of two nitrogen fixation regulatory regions, nifA and nfrX, in Azotobacter vinelandii and Azotobacter chroococcum. . Molecular Microbiology 2, 303-314.
    [Google Scholar]
  34. Santero, E.,, Hoover, T.,, Keener, J., & Kustu, S.. ( 1989;). In vitro activity of the nitrogen fixation regulatory protein NifA. . Proceedings of the National Academy of Sciences of the United States of America 86, 7346-7350.
    [Google Scholar]
  35. Souza, E. M.,, Funayama, S.,, Rigo, L., & Pedrosa, F. O.. ( 1991;). Cloning and characterization of the nifA gene from Herbaspirillum seropedicae. . Canadian Journal of Microbiology 37, 425-429.
    [Google Scholar]
  36. Staden, R.. ( 1982;). Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. . Nucleic Acids Research 10, 4731-4751.
    [Google Scholar]
  37. Staden, R.. ( 1984;). Graphic methods to determine the function of nucleic acid sequences. . Nucleic Acids Research 12, 521-538.
    [Google Scholar]
  38. Thöny, B.,, Anthamatten, D., & Hennecke, H.. ( 1989;). Dual control of the Bradyrhizobium japonicum symbiotic nitrogen fixation regulatory operon fixRnifA : analysis of cis- and trans-acting elements. . Journal of Bacteriology 171, 4162-4169.
    [Google Scholar]
  39. Toukdarian, A., & Kennedy, C.. ( 1986;). Regulation of nitrogen metabolism in Azotobacter vinelandii : isolation of ntr and glnA genes and construction of ntr mutants. . EM BO Journal 5, 399-407.
    [Google Scholar]
  40. Walker, J. E.,, Saraste, M.,, Runswick, M. J., & Gay, N. J.. ( 1982;). Distantly related sequences in the α and β subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. . EMBO Journal 1, 945-951.
    [Google Scholar]
  41. Yang, C. C., & Nash, H. A.. ( 1989;). The interaction of E. coli IHF protein with its specific binding sites. . Cell 57, 869-880.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-7-1511
Loading
/content/journal/micro/10.1099/00221287-137-7-1511
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