1887

Abstract

Summary: The cytochrome complex in the obligately aerobic diazotroph is an oxidase, which, has a low affinity for oxygen and is required for respiratory protection of nitrogenase. Mutations caused by insertion of Tn5-B20 upstream of the structural genes () for cytochrome result in over-expression of this oxidase and, for unexplained reasons, inability of the organism to grow microaerobically. Cloning and sequencing of this upstream region revealed a gene, The deduced amino acid sequence of CydR indicates that it is a new member of the Fnr class of regulators and that it represses expression. Refined mapping data for three insertions in are presented. The cloned gene complemented anaerobic growth of mutants and strongly enhanced expression of a fusion in an mutant.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-7-2197
1997-07-01
2021-07-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/7/mic-143-7-2197.html?itemId=/content/journal/micro/10.1099/00221287-143-7-2197&mimeType=html&fmt=ahah

References

  1. Anthamatten D., Scherb B., Hennecke H. 1992; Characterization of a fixLJ-regulated Bradyrhizobium japonicum gene sharing similarity with the Escherichia coli fnr and Rhizobium meliloti fixK genes. J Bacteriol 174:2111–2120
    [Google Scholar]
  2. Batut J., Daveran-Mingot M.-L., David M., Jacobs J., Garnerone A. M., Kahn D. 1989; fixK, a gene homologous with fnr and crp from Escherichia coli, regulates nitrogen fixation genes both positively and negatively in Rhizobium meliloti . EMBO J 8:1279–1286
    [Google Scholar]
  3. Becker S., Holighaus G., Gabrielczyk T., Unden G. 1996; O2as the regulatory signal for FNR-dependent gene regulation in Escherichia coli . J Bacteriol 178:4515–4521
    [Google Scholar]
  4. Brendel V., Trifonov E. N. 1984; A computer algorithm for testing potential prokaryotic terminators. Nucleic Acids Res 12:4411–4427
    [Google Scholar]
  5. Chang A. C. Y., Cohen S. N. 1978; Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156
    [Google Scholar]
  6. Church G. M., Gibert W. 1984; Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995
    [Google Scholar]
  7. Colonna-Romano A. S. W., SchlUter A., Boistard P., Puhler A., Priefer U. B. 1990; An Fnr-like protein encoded in Rhizobium leguminosarum biovar viciae shows structural and functional homology to Rhizobium meliloti FixK. Mol Gen Genet 223:138–147
    [Google Scholar]
  8. Cuypers H., Zumft W. G. 1993; Anaerobic control of denitrification in Pseudomonas stutzeri escapes mutagenesis of a fnr-like gene. J Bacteriol 175:7236–7246
    [Google Scholar]
  9. D’mello R. A., Hill S., Poole R. K. 1994a; Determination of the oxygen affinities of terminal oxidases in Azotobacter vinelandii using the deoxygenation of oxyleghaemoglobin and oxymyoglobin: cytochrome bd is a low-affinity oxidase. Microbiology 140:1395–1402
    [Google Scholar]
  10. D’mello R., Palmer S., Hill S., Poole R. K. 1994b; Thecytochrome bd terminal oxidase of Azotobacter vinelandii: low temperature photodissociation spectrophotometry reveals reactivity of cytochromes b595 and d with both carbon monoxide and oxygen. FEMS Microbiol Lett 121:115–120
    [Google Scholar]
  11. D’mello R., Hill S., Poole R. K. 1996; The cytochrome bd quinol oxidase in Escherichia coli has an extremely high oxygen affinity and two oxygen-binding haems: implications for regulation of activity in vivo by oxygen inhibition. Microbiology 142:755–763
    [Google Scholar]
  12. D’mello R., Purchase D., Poole R. K., Hill S. 1997; Expression and content of terminal oxidases in Azotobacter vinelandii grown with excess NH+ 4 are modulated by O2 supply. Microbiology 143:231–237
    [Google Scholar]
  13. Gil A., Kroll R. G., Poole R. K. 1992; The cytochrome composition of the meat-spoilage bacterium Brochothrix thermosphacta: identification of cytochrome a3- and d-type terminal oxidases under various conditions. Arch Microbiol 158:226–233
    [Google Scholar]
  14. Green J., Bennett B., Jordan P., Ralph E. T., Thomson A. J., Guest J. R. 1996; Reconstitution of the [4Fe-4S] cluster in FNR and demonstration of the aerobic-anaerobic transcription switch in vitro . Biochem J 316:887–892
    [Google Scholar]
  15. Guest J. R., Green J., Irvine A. S., Spiro S. 1996 The FNR modulon and FNR-regulated gene expression. . In Regulation of Gene Expression in Escherichia coli , pp. 318–342 . Edited by Lin E. C. C., Lynch A. S. Georgetown, TX: R. G. Landes Company;
    [Google Scholar]
  16. Haddock B. A., Jones C. W. 1977; Bacterial respiration. Bacteriol Rev 41:47–99
    [Google Scholar]
  17. Harley C. B., Reynolds R. P. 1987; Analysis of E. coli promoter sequences. Nucleic Acids Res 15:2343–2361
    [Google Scholar]
  18. Herrin D. L. 1988; Rapid reversible staining of Northern blots prior to hybridization. Biotechnique 6:196–198
    [Google Scholar]
  19. Hill S. 1992 Physiology of nitrogen fixation in free-living heterotrophs. . In Biological Nitrogen Fixation , pp. 86–133 . Edited by Stacey G., Burris R. H., Evans H. J. London: Chapman & Hall;
    [Google Scholar]
  20. Jacobson M. R., Brigle K. E., Bennett L. T., Setterquist R. A., Wilson M. S., Cash V. L., Beynon J., Newton,.W. E. & Dean D. R. 1989; Physical and genetic map of the major nif gene cluster from Azotobacter vinelandii . J Bacteriol 171:1017–1027
    [Google Scholar]
  21. Jones C. W., 8c Poole R. K. 1985; The analysis of cytochromes. Methods Microbiol 18:285–328
    [Google Scholar]
  22. Joshi M., Dikshit K. 1994; Oxygen dependent regulation of Vitreoscilla globin gene: evidence for positive regulation by Fnr. Biochem Biophys Res Commun 202:535–542
    [Google Scholar]
  23. Kahlow M. A., Zuberi T. M., Gennis R. B., 8t Loehr T. M. 1991; Identification of a ferryl intermediate of Escherichia coli cytochrome d terminal oxidase by resonance Raman spectroscopy. Biochemistry 30:11485–11489
    [Google Scholar]
  24. Kelly M. J. S., Poole R. K., Yates M. G., Kennedy C. 1990; Cloning and mutagenesis of genes encoding the cytochrome bd terminal oxidase complex in Azotobacter vinelandii: mutants deficient in the cytochrome d complex are unable to fix nitrogen in air. J Bacteriol 172:6010–6019
    [Google Scholar]
  25. Khoroshilova N., Beinert H., Kiley P. J. 1995; Association of a polynuclear iron-sulfur center with a mutant Fnr protein enhances DNA binding. Proc Natl Acad Sci USA 92:2499–2503
    [Google Scholar]
  26. Kolonay J. F., Moshiri F., Gennis R. B., Kaysser T. M., Maier R. J. 1994; Purification and characterization of the cytochrome bd complex from Azotobacter vinelandii: comparison to the complex from Escherichia coli . J Bacteriol 176:4177–4181
    [Google Scholar]
  27. Lazazzera B. A., Beinert H., Khoroshilova N., Kennedy M. C., Kiley P. J. 1996; DNA binding and dimerization of the Fe-S- containing FNR protein from Escherichia coli are regulated by oxygen. J Biol Chem 271:2762–2768
    [Google Scholar]
  28. Leung D., Van der Oost J., Kelly M. J. S., Saraste M., Hill S., Poole R. K. 1994; Mutagenesis of a gene encoding a cytochrome o-like terminal oxidase of Azotobacter vinelandii: a cytochrome o mutant is aero-tolerant during nitrogen fixation. FEMS Microbiol Lett 119:351–358
    [Google Scholar]
  29. Li J., Stewart V. 1992; Localization of upstream sequence elements required for nitrate and anaerobic induction of fdn (formate dehydrogenase-N) operon expression in Escherichia coli K-12. J Bacteriol 174:4935–4942
    [Google Scholar]
  30. MacInnes J. I., Kim J. E., Lian C. J., Soltes G. A. 1990; Actinobacillus pleuropneumoniae hlyX gene homology with the fnr gene of Escherichia coli. J Bacteriol 172:4587–4592
    [Google Scholar]
  31. Markwell M. A. K., Haas S. M., Bieber L., L.& Tolbert N. E. 1978; A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87:206–210
    [Google Scholar]
  32. Melville S. B., Gunsalus R. P. 1990; Mutations in fnr that alter anaerobic regulation of electron transport-associated genes in Escherichia coli . J Biol Chem 265:18733–18736
    [Google Scholar]
  33. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  34. Moshiri F., Chawla A., 8t Maier R. J. 1991a; Cloning, characterization and expression in Escherichia coli of the gene encoding the cytochrome d oxidase complex from Azotobacter vinelandii . J Bacteriol 173:6230–6241
    [Google Scholar]
  35. Moshiri F., Smith E. G., Taormino J. P., Maier R. J. 1991b; Transcriptional regulation of cytochrome d in nitrogen-fixing Azotobacter vinelandii. Evidence that up-regulation during N2 fixation is independent of nifA but dependent on ntrA . J Biol Chem 266:23169–23174
    [Google Scholar]
  36. Murphy G., Ward E. S. 1989 Sequencing of double-stranded DNA. . In Nucleic Acids Sequencing: a Practical Approach , pp. 99–115 . Edited by Howe C. J., Ward E. S. Oxford: IRL Press;
    [Google Scholar]
  37. Patschkowski T., Schluter A., Priefer U. B. 1996; Rhizobium leguminosarum bv viciae contains a second fnr/fixK-like gene and an unusual fixL homologue. Mol Microbiol 21:267–280
    [Google Scholar]
  38. Poole R. K. 1994; Oxygen reactions with bacterial oxidases and globins: binding, reduction and regulation. Antonie Leeuwenhoek 65:289–310
    [Google Scholar]
  39. Poole R. K., Hill S. 1997; Respiratory protection of nitrogenase activity in Azotobacter vinelandii - roles of the terminal oxidases. Biosci Rep (in press)
    [Google Scholar]
  40. Poole R. K., Williams H. D. 1988; Formation of the 680 nm-absorbing form of the cytochrome bd oxidase complex of Escherichia coli by reaction of hydrogen peroxide with the ferric form. FEBS Lett 231:243–246
    [Google Scholar]
  41. Robson R. L., Postgate J. R. 1980; Oxygen and hydrogen in biological nitrogen fixation. Annu Rev Microbiol 34:183–207
    [Google Scholar]
  42. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor; NY: Cold Spring Harbor Laboratory:
    [Google Scholar]
  43. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  44. Sawers G. 1991; Identification and molecular characterization of a transcriptional regulator from Pseudomonas aeruginosa PAOl exhibiting structural and functional similarity to the Fnr protein of Escherichia coli . Mol Microbiol 5:1469–1481
    [Google Scholar]
  45. Sawers R. G., Zehelein E., Bfick A. 1988; Two-dimensional gel electrophoretic analysis of Escherichia coli proteins: influence of various anaerobic growth conditions and the fnr gene product on cellular protein composition. Arch Microbiol 149:240–244
    [Google Scholar]
  46. Schlüiter A., Patschkowski T., Unden G., Priefer U. B. 1992; The Rhizobium leguminosarum FnrN protein is functionally similar to Escherichia coli Fnr and promotes heterologous oxygen-dependent activation of transcription. Mol Microbiol 6:3395–3404
    [Google Scholar]
  47. Sharrocks A. D., Green J., Guest J. R. 1990; In vivo and in vitro mutants of Fnr, the anaerobic transcriptional regulator of Escherichia coli . FEBS Lett 270:119–122
    [Google Scholar]
  48. Shaw D. J., Guest J. R. 1982; Nucleotide sequence of the fnr gene and primary structure of the Fnr protein of Escherichia coli . Nucleic Acids Res 10:6119–6130
    [Google Scholar]
  49. Silhavy T. J, ., Berman M. L., Enquist L. W. 1984 Experiments with Gene Fusions , pp. 102–112 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  50. Spiro S., Guest J. R. 1987; Activation of the lac operon of Escherichia coli by a mutant Fnr protein. Mol Microbiol 1:53–58
    [Google Scholar]
  51. Spiro S., Guest J. R. 1988; Inactivation of the Fnr protein of Escherichia coli by targeted mutagenesis in the N-terminal region. Mol Microbiol 2:701–707
    [Google Scholar]
  52. Stewart V. 1982; Requirement of Fnr and NarL functions for nitrate reductase expression in Escherichia coli K-12. J Bacteriol 151:1320–1325
    [Google Scholar]
  53. Stewart V., Parales J. , Jr 1988; Identification and expression of genes narL and narX of the nar (nitrate reductase) locus in Escherichia coli K-12. J Bacteriol 170:1589–1597
    [Google Scholar]
  54. Thӧny-Meyer L., Beck C., Preisig O., Hennecke H. 1994; The ccoNOQP gene cluster codes for a cb-type cytochrome oxidase that functions in aerobic respiration of Rhodobacter capsulatus . Mol Microbiol 14:705–716
    [Google Scholar]
  55. Wang R. F., Kushner S. R. 1991; Construction of versatile low- copy-number vector for cloning, sequencing and gene expression in Escherichia coli . Gene 100:195–199
    [Google Scholar]
  56. Williams H. D., Poole R. K. 1987; The cytochromes of Acetobacter pasteurianus NCIB 6428. Evidence of a role for a cytochrome a1-like haemoprotein in electron transfer to cytochrome oxidase d . J Gen Microbiol 133:2461–2472
    [Google Scholar]
  57. Wu G., Williams H. D., Zamanian M., Gibson F., Poole R. K. 1992; Isolation and characterization of Escherichia coli mutants affected in aerobic respiration: the cloning and nucleotide sequence of ubiG. Identification of an S-adenosylmethionine- binding motif in protein, RNA, and small-molecule methyl- transferases. J Gen Microbiol 138:2101–2112
    [Google Scholar]
  58. Wu G., Williams H. D., Gibson F., Poole R. K. 1993; Mutants of Escherichia coli affected in respiration: the cloning and nucleotide sequence of ubiA, encoding the membrane-bound p- hydroxybenzoate: octaprenyltransferase. J Gen Microbiol 139:1795–1805
    [Google Scholar]
  59. Yang T. 1986; Biochemical and biophysical properties of cytochrome o of Azotobacter vinelandii . Biochim Biophys Acta 848:342–351
    [Google Scholar]
  60. Zimmermann A., Reimmann C., Galimanel M., Haas D. 1991; Anaerobic growth and cyanide synthesis of Pseudomonas aeruginosa dependent on Anr, a regulatory gene homologous with Fnr of Escherichia coli . Mol Microbiol 15:1483–1490
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-7-2197
Loading
/content/journal/micro/10.1099/00221287-143-7-2197
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