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. 1995 Apr;177(8):2214–2217. doi: 10.1128/jb.177.8.2214-2217.1995

Bradyrhizobium japonicum cytochrome c550 is required for nitrate respiration but not for symbiotic nitrogen fixation.

M Bott 1, L Thöny-Meyer 1, H Loferer 1, S Rossbach 1, R E Tully 1, D Keister 1, C A Appleby 1, H Hennecke 1
PMCID: PMC176869  PMID: 7721713

Abstract

Bradyrhizobium japonicum possesses three soluble c-type cytochromes, c550, c552, and c555. The genes for cytochromes c552 (cycB) and c555 (cycC) were characterized previously. Here we report the cloning, sequencing, and mutational analysis of the cytochrome c550 gene (cycA). A B. japonicum mutant with an insertion in cycA failed to synthesize a 12-kDa c-type cytochrome. This protein was detectable in the cycA mutant complemented with cloned cycA, which proves that it is the cycA gene product. The cycA mutant, a cycB-cycC double mutant, and a cycA-cycB-cycC triple mutant elicited N2-fixing root nodules on soybean (Nod+ Fix+ phenotype); hence, none of these three cytochromes c is essential for respiration supporting symbiotic N2 fixation. However, cytochrome c550, in contrast to cytochromes c552 and c555, was shown to be essential for anaerobic growth of B. japonicum, using nitrate as the terminal electron acceptor.

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Selected References

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  1. Ambler R. P., Daniel M., Hermoso J., Meyer T. E., Bartsch R. G., Kamen M. D. Cytochrome c2 sequence variation among the recognised species of purple nonsulphur photosynthetic bacteria. Nature. 1979 Apr 12;278(5705):659–660. doi: 10.1038/278659a0. [DOI] [PubMed] [Google Scholar]
  2. Appleby C. A. Electron transport systems of Rhizobium japonicum. I. Haemoprotein P-450, other CO-reactive pigments, cytochromes and oxidases in bacteroids from N2-fixing root nodules. Biochim Biophys Acta. 1969 Jan 14;172(1):71–87. doi: 10.1016/0005-2728(69)90093-0. [DOI] [PubMed] [Google Scholar]
  3. Appleby C. A., James P., Hennecke H. Characterization of three soluble c-type cytochromes isolated from soybean root nodule bacteroids of Bradyrhizobium japonicum strain CC705. FEMS Microbiol Lett. 1991 Oct 1;67(2):137–144. doi: 10.1016/0378-1097(91)90344-a. [DOI] [PubMed] [Google Scholar]
  4. Bott M., Ritz D., Hennecke H. The Bradyrhizobium japonicum cycM gene encodes a membrane-anchored homolog of mitochondrial cytochrome c. J Bacteriol. 1991 Nov;173(21):6766–6772. doi: 10.1128/jb.173.21.6766-6772.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Daniel R. M., Appleby C. A. Anaerobic-nitrate, symbiotic and aerobic growth of Rhizobium japonicum: effects on cytochrome P 450 , other haemoproteins, nitrate and nitrite reductases. Biochim Biophys Acta. 1972 Sep 20;275(3):347–354. doi: 10.1016/0005-2728(72)90215-0. [DOI] [PubMed] [Google Scholar]
  6. De Vos G. F., Walker G. C., Signer E. R. Genetic manipulations in Rhizobium meliloti utilizing two new transposon Tn5 derivatives. Mol Gen Genet. 1986 Sep;204(3):485–491. doi: 10.1007/BF00331029. [DOI] [PubMed] [Google Scholar]
  7. Francis R. T., Jr, Becker R. R. Specific indication of hemoproteins in polyacrylamide gels using a double-staining process. Anal Biochem. 1984 Feb;136(2):509–514. doi: 10.1016/0003-2697(84)90253-7. [DOI] [PubMed] [Google Scholar]
  8. Grisshammer R., Wiessner C., Michel H. Sequence analysis and transcriptional organization of the Rhodopseudomonas viridis cytochrome c2 gene. J Bacteriol. 1990 Sep;172(9):5071–5078. doi: 10.1128/jb.172.9.5071-5078.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Keister D. L., Marsh S. S. Hemoproteins of Bradyrhizobium japonicum Cultured Cells and Bacteroids. Appl Environ Microbiol. 1990 Sep;56(9):2736–2741. doi: 10.1128/aem.56.9.2736-2741.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Preisig O., Anthamatten D., Hennecke H. Genes for a microaerobically induced oxidase complex in Bradyrhizobium japonicum are essential for a nitrogen-fixing endosymbiosis. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3309–3313. doi: 10.1073/pnas.90.8.3309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ramseier T. M., Göttfert M. Codon usage and G + C content in Bradyrhizobium japonicum genes are not uniform. Arch Microbiol. 1991;156(4):270–276. doi: 10.1007/BF00262997. [DOI] [PubMed] [Google Scholar]
  12. Ramseier T. M., Winteler H. V., Hennecke H. Discovery and sequence analysis of bacterial genes involved in the biogenesis of c-type cytochromes. J Biol Chem. 1991 Apr 25;266(12):7793–7803. [PubMed] [Google Scholar]
  13. Ritz D., Bott M., Hennecke H. Formation of several bacterial c-type cytochromes requires a novel membrane-anchored protein that faces the periplasm. Mol Microbiol. 1993 Aug;9(4):729–740. doi: 10.1111/j.1365-2958.1993.tb01733.x. [DOI] [PubMed] [Google Scholar]
  14. Rossbach S., Loferer H., Acuña G., Appleby C. A., Hennecke H. Cloning, sequencing and mutational analysis of the cytochrome c552 gene (cycB) from Bradyrhizobium japonicum strain 110. FEMS Microbiol Lett. 1991 Oct 1;67(2):145–152. doi: 10.1016/0378-1097(91)90345-b. [DOI] [PubMed] [Google Scholar]
  15. Tanaka Y., Fukumori Y., Yamanaka T. The complete amino acid sequence of Nitrobacter agilis cytochrome c-550. Biochim Biophys Acta. 1982 Sep 22;707(1):14–20. doi: 10.1016/0167-4838(82)90390-9. [DOI] [PubMed] [Google Scholar]
  16. Thöny-Meyer L., Stax D., Hennecke H. An unusual gene cluster for the cytochrome bc1 complex in Bradyrhizobium japonicum and its requirement for effective root nodule symbiosis. Cell. 1989 May 19;57(4):683–697. doi: 10.1016/0092-8674(89)90137-2. [DOI] [PubMed] [Google Scholar]
  17. Tully R. E., Sadowsky M. J., Keister D. L. Characterization of cytochromes c550 and c555 from Bradyrhizobium japonicum: cloning, mutagenesis, and sequencing of the c555 gene (cycC). J Bacteriol. 1991 Dec;173(24):7887–7895. doi: 10.1128/jb.173.24.7887-7895.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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