• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. May 1994; 176(9): 2560–2568.
PMCID: PMC205393

Functional analysis of the fixNOQP region of Azorhizobium caulinodans.

Abstract

The deduced amino acid sequences of four open reading frames identified upstream of the fixGHI region in Azorhizobium caulinodans are very similar to the putative terminal oxidase complex coded by the fixNOQP operons from Rhizobium meliloti and Bradyrhizobium japonicum. The expression of the A. caulinodans fixNOQP genes, which was maximal under microaerobiosis, was positively regulated by FixK and independent of NifA. In contrast to the Fix- phenotype of B. japonicum and R. meliloti fixN mutants, an A. caulinodans fixNO-deleted mutant strain retained 50% of the nitrogenase activity of the wild type in the symbiotic state. In addition, the nitrogenase activity was scarcely reduced under free-living conditions. Analysis of membrane fractions of A. caulinodans wild-type and mutant strains suggests that the fixNOQP region encodes two proteins with covalently bound hemes, tentatively assigned to fixO and fixP. Spectral analysis showed a large decrease in the c-type cytochrome content of the fixN mutant compared with the wild type. These results provide evidence for the involvement of FixNOQP proteins in a respiratory process. The partial impairment in nitrogen fixation of the fixN mutant in planta may be due to the activity of an alternative terminal oxidase compensating for the loss of the oxidase complex encoded by fixNOQP.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allen LN, Hanson RS. Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol. J Bacteriol. 1985 Mar;161(3):955–962. [PMC free article] [PubMed]
  • Batut J, Daveran-Mingot ML, David M, Jacobs J, Garnerone AM, Kahn D. fixK, a gene homologous with fnr and crp from Escherichia coli, regulates nitrogen fixation genes both positively and negatively in Rhizobium meliloti. EMBO J. 1989 Apr;8(4):1279–1286. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • David M, Daveran ML, Batut J, Dedieu A, Domergue O, Ghai J, Hertig C, Boistard P, Kahn D. Cascade regulation of nif gene expression in Rhizobium meliloti. Cell. 1988 Aug 26;54(5):671–683. [PubMed]
  • Dreyfus BL, Elmerich C, Dommergues YR. Free-living Rhizobium strain able to grow on n(2) as the sole nitrogen source. Appl Environ Microbiol. 1983 Feb;45(2):711–713. [PMC free article] [PubMed]
  • Elmerich C, Dreyfus BL, Reysset G, Aubert JP. Genetic analysis of nitrogen fixation in a tropical fast-growing Rhizobium. EMBO J. 1982;1(4):499–503. [PMC free article] [PubMed]
  • Francis RT, Jr, Becker RR. Specific indication of hemoproteins in polyacrylamide gels using a double-staining process. Anal Biochem. 1984 Feb;136(2):509–514. [PubMed]
  • Hill S, Viollet S, Smith AT, Anthony C. Roles for enteric d-type cytochrome oxidase in N2 fixation and microaerobiosis. J Bacteriol. 1990 Apr;172(4):2071–2078. [PMC free article] [PubMed]
  • Kahn D, David M, Domergue O, Daveran ML, Ghai J, Hirsch PR, Batut J. Rhizobium meliloti fixGHI sequence predicts involvement of a specific cation pump in symbiotic nitrogen fixation. J Bacteriol. 1989 Feb;171(2):929–939. [PMC free article] [PubMed]
  • Kaminski PA, Elmerich C. Involvement of fixLJ in the regulation of nitrogen fixation in Azorhizobium caulinodans. Mol Microbiol. 1991 Mar;5(3):665–673. [PubMed]
  • Kaminski PA, Mandon K, Arigoni F, Desnoues N, Elmerich C. Regulation of nitrogen fixation in Azorhizobium caulinodans: identification of a fixK-like gene, a positive regulator of nifA. Mol Microbiol. 1991 Aug;5(8):1983–1991. [PubMed]
  • Kelly MJ, Poole RK, Yates MG, Kennedy C. 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. 1990 Oct;172(10):6010–6019. [PMC free article] [PubMed]
  • Kitts CL, Ludwig RA. Azorhizobium caulinodans respires with at least four terminal oxidases. J Bacteriol. 1994 Feb;176(3):886–895. [PMC free article] [PubMed]
  • Kokotek W, Lotz W. Construction of a lacZ-kanamycin-resistance cassette, useful for site-directed mutagenesis and as a promoter probe. Gene. 1989 Dec 14;84(2):467–471. [PubMed]
  • Labbe P, Chaix P. Nouvelle technique de réalisation de spectres d'absorption à basse température. Bull Soc Chim Biol (Paris) 1969;51(12):1642–1644. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lemieux LJ, Calhoun MW, Thomas JW, Ingledew WJ, Gennis RB. Determination of the ligands of the low spin heme of the cytochrome o ubiquinol oxidase complex using site-directed mutagenesis. J Biol Chem. 1992 Jan 25;267(3):2105–2113. [PubMed]
  • Mandon K, Kaminski PA, Mougel C, Desnoues N, Dreyfus B, Elmerich C. Role of the fixGHI region of Azorhizobium caulinodans in free-living and symbiotic nitrogen fixation. FEMS Microbiol Lett. 1993 Dec 1;114(2):185–189. [PubMed]
  • Minagawa J, Mogi T, Gennis RB, Anraku Y. Identification of heme and copper ligands in subunit I of the cytochrome bo complex in Escherichia coli. J Biol Chem. 1992 Jan 25;267(3):2096–2104. [PubMed]
  • Neidle EL, Kaplan S. Rhodobacter sphaeroides rdxA, a homolog of Rhizobium meliloti fixG, encodes a membrane protein which may bind cytoplasmic [4Fe-4S] clusters. J Bacteriol. 1992 Oct;174(20):6444–6454. [PMC free article] [PubMed]
  • O'Brian MR, Maier RJ. Molecular aspects of the energetics of nitrogen fixation in Rhizobium-legume symbioses. Biochim Biophys Acta. 1989 May 30;974(3):229–246. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Saraste M. Structural features of cytochrome oxidase. Q Rev Biophys. 1990 Nov;23(4):331–366. [PubMed]
  • Smith A, Hill S, Anthony C. The purification, characterization and role of the d-type cytochrome oxidase of Klebsiella pneumoniae during nitrogen fixation. J Gen Microbiol. 1990 Jan;136(1):171–180. [PubMed]
  • Soberón M, Membrillo-Hernández J, Aguilar GR, Sánchez F. Isolation of Rhizobium phaseoli Tn5-induced mutants with altered expression of cytochrome terminal oxidases o and aa3. J Bacteriol. 1990 Mar;172(3):1676–1680. [PMC free article] [PubMed]
  • Soberón M, Williams HD, Poole RK, Escamilla E. Isolation of a Rhizobium phaseoli cytochrome mutant with enhanced respiration and symbiotic nitrogen fixation. J Bacteriol. 1989 Jan;171(1):465–472. [PMC free article] [PubMed]
  • Spiro S, Guest JR. FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev. 1990 Aug;6(4):399–428. [PubMed]
  • Stigter J, Schneider M, de Bruijn FJ. Azorhizobium caulinodans nitrogen fixation (nif/fix) gene regulation: mutagenesis of the nifA -24/-12 promoter element, characterization of a ntrA(rpoN) gene, and derivation of a model. Mol Plant Microbe Interact. 1993 Mar-Apr;6(2):238–252. [PubMed]
  • 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. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...