1887

Microbiology Society logo

Volume 142, Issue 12

The cytochrome complex but not CycM is necessary for symbiotic nitrogen fixation by Free

Abstract

Following Tn5 mutagenesis of biovar two mutants in one complementation group were identified as being unable to fix nitrogen in pea nodules. Spectroscopic analysis revealed that the mutants had lowered levels of -type cytochromes and cytochromes , but increased levels of cytochrome Cells of the mutants were greatly reduced in their ability to oxidize the artificial electron donor -tetramethyl--phenylenediamine but membranes prepared from them had increased levels of succinate-and NADH-dependent respiration. NADH oxidation by the mutants was insensitive to the respiratory inhibitor antimycin A, that targets the cytochrome complex. Molecular analysis of the mutants revealed that they were affected in the cytochrome complex. One of the mutants contained Tn in a gene homologous to that encoding cytochrome , and in the other the Tn was in DNA homologous to that encoding the cytochrome component of the cytochrome complex. Haem staining revealed that haem proteins of 31000 and 23000 were absent from membranes from the mutants whereas an additional soluble -type cytochrome protein of 23000 was present. We conclude that the larger of these two haem proteins corresponds to cytochrome and, in its absence, the protein of 23000 does not remain associated with the membrane. Formation of this 23000 component was specifically blocked in a third respiratory-defective mutant which contained Tn in a region of DNA showing homology to a gene previously shown to encode the membrane-bound -type cytochrome CycM. Although the cytochrome complex is essential for symbiotic nitrogen fixation, the other membrane-bound -type cytochrome (CycM) is not.

  • Published Online:
Keyword(s): bacteroids , cytochromes , Keywords: Rhizobium, , nitrogen fixation and symbiosis
© Society for General Microbiology 1996
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-12-3381
1996-12-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/12/mic-142-12-3381.html?itemId=/content/journal/micro/10.1099/13500872-142-12-3381&mimeType=html&fmt=ahah

References

  1. Appleby C. A. 1969; Electron transport systems of Rhizobium japonicum. II. Rhizobium haemoglobin, cytochromes and oxidases in free-living (cultured) cells. Biochim Biophys Acta 172:88–105
     [Google Scholar]
  2. Appleby C. A. 1984; Leghemoglobin and Rhizobium respiration. Annu Rep Plant Physiol 35:443–478
     [Google Scholar]
  3. Beringer J. E. 1974; R factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198
     [Google Scholar]
  4. Bott M., Bolliger M., Hennecke H. 1990; Genetic analysis of the cytochrome c-aa3 branch of the Bradyrhizobium japonicumrespiratory chain. Mol Microbiol 4:2147–2157
     [Google Scholar]
  5. Bott M., Ritz D., Hennecke H. 1991; The Bradyrhizobium japonicum cycM gene encodes a membrane-anchored homolog of mitochondrial cytochrome c. J Bacteriol 173:6766–6772
     [Google Scholar]
  6. Buchanan-Wollaston A. V. 1979; Generalized transduction in Rhizobium leguminosarum. J Gen Microbiol 112:135–142
     [Google Scholar]
  7. Delgado M. -J., Yeoman K. H., Wu G., Vargas C., Davies A. E., Poole R. K., Johnston A. W. B., Downie J. A. 1995; Characterization of the cycHJKL genes involved in cytochrome cbiogenesis and symbiotic nitrogen fixation in Rhizobium leguminosarum. J Bacteriol 177:4927–4934
     [Google Scholar]
  8. D’mello R., Palmer S., Hill S., Poole R. K. 1994; The cytochrome 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]
  9. Downie J. A., Hombrecher G., Ma Q. -S., Knight C. D., Wells B., Johnston A. W. B. 1983; Cloned nodulation genes of Rhizobium leguminosarum determine host-range specificity. Mol Gen Genet 190:359–365
     [Google Scholar]
  10. Figurski D. H., Helinski D. R. 1979; Replication of an origin- containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 761648–1652
     [Google Scholar]
  11. Gabellini N., Sebald W. 1986; Nucleotide sequence and transcription of the fbc operon from Rhodopseudomonas sphaeroides:evaluation of the deduced amino acid sequences of the FeS protein, cytochrome b and cytochrome c1. Eur J Biochem 154:569–579
     [Google Scholar]
  12. Greenwood G., Wilson M. T., Brunori M. 1974; Studies on partially reduced mammalian cytochrome oxidase; reaction with carbon monoxide and oxygen. Biochem J 137:205–215
     [Google Scholar]
  13. Kahn D., Batut J., Daveran M. L., Fourment J. 1993; Structure and regulation of the fixNOQP operon from Rhizobium meliloti. In New Horizons in Nitrogen Fixation p. 474 Edited by Palacios R., Mora J., Newton W. E. Dordrecht: Kluwer;
     [Google Scholar]
  14. Kitts C. L., Ludwig R. A. 1994; Azorhizobium caulinodans respires with at least four terminal oxidases. J Bacteriol 176:886–895
     [Google Scholar]
  15. Knight C. D., Rossen L., Robertson J. G., Wells B., Downie J. A. 1986; Nodulation inhibition by Rhizobium leguminosarummulticopy nodABC genes and analysis of early stages of plant infection. J Bacteriol 166:552–558
     [Google Scholar]
  16. Kretovich W. L., Romanov V. I., Korolyov A. V. 1973; Rhizobium leguminosarum cytochromes (Vicia faba). Plant Soil 39:619–634
     [Google Scholar]
  17. Lamb J. W., Hombrecher G., Johnston A. W. B. 1982; Plasmid- determined nodulation and nitrogen-fixation abilities in Rhispbium phaseoli. Mol Gen Genet 186:449–452
     [Google Scholar]
  18. Mandon K., Kaminski P. A., Elmerich C. 1994; Functional analysis of the fixNOQP region of Azorhizobium caulinodans. J Bacteriol 176:2560–2568
     [Google Scholar]
  19. Marrs B., Gest H. 1973; Genetic mutations affecting the respiratory electron-transport system of the photosynthetic bacterium Rhodopseudomonas capsulata. J Bacteriol 114:1045–1051
     [Google Scholar]
  20. O’Brian M. R., Maier R. J. 1989; Molecular aspects of the energetics of nitrogen fixation in Rhizobium-legume symbioses. Biochim Biophys Acta 974:229–246
     [Google Scholar]
  21. Poole R. K. 1994; Oxygen reactions with bacterial oxidases and globins: binding, reduction and regulation. Antonie Leeuwenhoek 65:289–310
     [Google Scholar]
  22. Poole R. K., Lloyd D., Chance B. 1979; The reaction of cytochrome oxidase with oxygen in the fission yeast Schizo-saccharomycespombe 972h−. Biochem J 184:555–563
     [Google Scholar]
  23. Preisig O., Anthamatten D., Hennecke H. 1993; Genes for a microaerobically induced oxidase complex in Bradyrhizobium japonicum are essential for a nitrogen-fixing endosymbiosis. Proc Natl Acad Sci USA 903309–3313
     [Google Scholar]
  24. 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]
  25. Schulter A., Patschkowski T., Weidner S., Unden G., Hynes M. F., Priefer U. B. 1993; Functional and regulatory characteristics of FnrN, an oxygen-responsive transcriptional activator in Rhizobium leguminosarum bv.viciae. In New Horizons in Nitrogen Fixation p. 493 Edited by Palacios R., Mora J., Newton W. E. Dordrecht: Kluwer;
     [Google Scholar]
  26. Simon R., Priefer U., Pühler A. 1983; A broad host-range mobilization system for in vivo genetic engineering: transposon mutagenesis of Gram-negative bacteria. Biotechnol 1:784–791
     [Google Scholar]
  27. Thöny-Meyer L., Stax D., Hennecke H. 1989; An unusual gene cluster for the cytochrome bc1 complex in Bradyrhizobium japonicumand its requirement for effective root nodule symbiosis. Cell 57:683–697
     [Google Scholar]
  28. Thöny-Meyer L., James P., Hennecke H. 1991; From one gene to two proteins: the biogenesis of cytochromes b and c1 in Bradyrhizobium japonicum. Proc Natl Acad Sci USA 885001–5005
     [Google Scholar]
  29. Vargas C., McEwan A. G., Downie J. A. 1993; Detection of c-type cytochromes using enhanced chemiluminescence. Anal Biochem 209:323–326
     [Google Scholar]
  30. Vargas C., Wu G., Davies A. E., Downie J. A. 1994; Identification of a gene encoding a thioredoxin-like product necessary for cytochrome c biosynthesis and symbiotic nitrogen fixation in Rhizobium leguminosarum. J Bacteriol 176:4117–4123
     [Google Scholar]
  31. Vargas C., Guanghui W., Delgado M. -J., Poole R. K., Downie J. A. 1996; Identification of symbiosis-specific c-type cytochromes and a putative oxidase in bacteroids of Rhizobium leguminosarumbiovar viciae. Microbiol 142:41–46
     [Google Scholar]
  32. Wikström M., Krab K., Saraste M. 1981 Cytochrome Oxidase: a Synthesis London: Academic Press;
     [Google Scholar]
  33. Williams H. D., Appleby C. A., Poole R. K. 1990; The unusual behaviour of the putative terminal oxidases of Bradyrhizobium japonicum bacteroids revealed by low-temperature photodissociation studies. Biochim Biophys Acta 1019:225–232
     [Google Scholar]
  34. 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]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-12-3381
Loading
The cytochrome bc1 complex but not CycM is necessary for symbiotic nitrogen fixation by Rhizobium leguminosarum
Microbiology 142, 3381 (1996); https://doi.org/10.1099/13500872-142-12-3381
/content/journal/micro/10.1099/13500872-142-12-3381
/content/journal/micro/10.1099/13500872-142-12-3381
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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