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A novel response-regulator is able to suppress the nodulation defect of a Bradyrhizobium japonicum nodW mutant

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Abstract

The two-component regulatory system Nod-VW of Bradyrhizobium japonicum is essential for the nodulation of the legume host plants Vigna radiata, V. unguiculata and Macroptilium atropurpureum. The NodV protein shares homology with the sensor-kinases, whereas the NodW protein is a member of the response-regulator class. We report here the identification of a new B. japonicum DNA region that is able to suppress the phenotypic defect of a nodW mutant, provided that this region is expressed from a foreign promoter. The minimal complementing region, which itself is not essential for nodulation in a nodW + background, consists of one gene designated nwsB (nodW-suppressor). The deduced amino acid sequence of the nwsB gene product shows a high degree of homology to NodW. The nwsB gene is preceded by a long open reading frame, nwsA, whose putative product appears to be a sensor-kinase. Downstream of nwsB, an open reading frame encoding a second putative response-regulator was identified. Interspecies hybridization revealed the presence of nwsAB-like DNA also in other Bradyrhizobium strains. Using nwsB′-′lacZ fusions, the nwsB gene was found to be expressed rather weakly in B. japonicum. This low level of expression is obviously not sufficient to compensate for a nodW defect, whereas strong overexpression of nwsB is a condition that leads to suppression of the nodW mutation.

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References

  • Albright LM, Huala E, Ausubel FM (1989) Prokaryotic signal transduction mediated by sensor and regulator protein pairs. Annu Rev Genet 23:311–336

    Google Scholar 

  • Anthamatten D, Hennecke H (1991) The regulatory status of the fixL- and fixJ-like genes in Bradyrhizobium japonicum may be different from that in Rhizobium meliloti. Mol Gen Genet 225:38–48

    Google Scholar 

  • 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 

  • Beringer JE, Brewin NJ, Johnston AWB (1980) The genetic analysis of Rhizobium in relation to symbiotic nitrogen fixation. Heredity 45:161–186

    Google Scholar 

  • Bott M, Bolliger M, Hennecke H (1990) Genetic analysis of the cytochrome c-aa 3 branch of the Bradyrhizobium japonicum respiratory chain. Mol Microbiol 4:2147–2157

    Google Scholar 

  • Chiang RC, Cavicchioli R, Gunsalus RP (1992) Identification and characterization of narQ, a second nitrate sensor for nitrate-dependent gene regulation in Escherichia coli. Mol Microbiol 6:1913–1923

    Google Scholar 

  • Corbin D, Barran L, Ditta G (1983) Organization and expression of Rhizobium meliloti nitrogen fixation genes. Proc Natl Acad Sci USA 80:3005–3009

    Google Scholar 

  • Davis RW, Botstein D, Roth JR (1980) Advanced bacterial genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Dénarié J, Debellé F, Rosenberg C (1992) Signaling and host range variation in nodulation. Annu Rev Microbiol 46:497–531

    Google Scholar 

  • Dreyfus B, Garcia JL, Gillis M (1988) Characterization of Azorhizobium caulinodans gen. nov. sp. nov., a stem-nodulating nitrogen-fixing bacterium isolated from Sesbania rostrata. Int J Syst Bacteriol 38:89–98

    Google Scholar 

  • Figurski DH, Helinski DR (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652

    Google Scholar 

  • Fisher RF, Long SR (1992) Rhizobium-plant signal exchange. Nature 357:655–660

    Google Scholar 

  • Göttfert M (1993) Regulation and function of rhizobial nodulation genes. FEMS Microbiol Rev 104:39–64

    Google Scholar 

  • Göttfert M, Lamb JW, Gasser R, Semenza J, Hennecke H (1989) Mutational analysis of the Bradyrhizobium japonicum common nod genes and further nod box-linked genomic DNA regions. Mol Gen Genet 215:407–415

    Google Scholar 

  • Göttfert M, Grob P, Hennecke H (1990a) Proposed regulatory pathway encoded by the nodV and nodW genes, determinants of host specificity in Bradyrhizobium japonicum. Proc Natl Acad Sci USA 87:2680–2684

    Google Scholar 

  • Göttfert M, Hitz S, Hennecke H (1990b) Identification of nodS and nodU, two inducible genes inserted between the Bradyrhizobium japonicum nodYABC and nodIJ genes. Mol Plant-Microbe Interact 3:308–316

    Google Scholar 

  • Göttfert M, Holzhäuser D, Bäni D, Hennecke H (1992) Structural and functional analysis of two different nodD genes in Bradyrhizobium japonicum USDAl10. Mol Plant-Microbe Interact 5:257–265

    Google Scholar 

  • Gross R, Arico B, Rappuoli R (1989) Families of bacterial signal-transducing proteins. Mol Microbiol 3:1661–1667

    Google Scholar 

  • Györgypal Z, Kiss GB, Kondorosi A (1991) Transduction of plant signal molecules by the Rhizobium NodD proteins. Bioessays 13:575–581

    Google Scholar 

  • Hahn M, Hennecke H (1984) Localized mutagenesis in Rhizobium japonicum. Mol Gen Genet 193:46–52

    Google Scholar 

  • Hanahan D (1983) Studies of transformation of Escherichia coli with plasmids. J Mol Biol 166:557–563

    Google Scholar 

  • Hennecke H, Günther I, Binder F (1982) A novel cloning vector for the direct selection of recombinant DNA in E. coli. Gene 19:231–234

    Google Scholar 

  • Igo MM, Ninfa AJ, Stock JB, Silhavy TJ (1989) Phosphorylation and dephosphorylation of a bacterial transcriptional activator by a transmembrane receptor. Genes Dev 3:1725–1734

    Google Scholar 

  • Kondorosi A, Kondorosi E, John M, Schmidt J, Schell J (1991) The role of nodulation genes in bacterium-plant communication. Genet Eng 13:115–136

    Google Scholar 

  • Kullik I, Fritsche S, Knobel H, Sanjuan J, Hennecke H, Fischer H-M (1991) Bradyrhizobium japonicum has two differentially regulated, functional homologs of the σ54 gene (rpoN). J Bacteriol 173:1125–1138

    Google Scholar 

  • Lamb JW, Hombrecher G, Johnston AWB (1982) Plasmid-determined nodulation and nitrogen fixation abilities in Rhizobium phaseoli. Mol Gen Genet 186:449–452

    Google Scholar 

  • Lerouge P, Roche P, Faucher C, Maillet F, Truchet G, Promé JC, Dénarié J (1990) Symbiotic host-specificity of Rhizobium meliloti is determined by a sulphated and acylated glucosamine oligosaccharide signal. Nature 344:781–784

    Google Scholar 

  • Long SR (1992) Genetic analysis of Rhizobium nodulation. In: Stacey G, Burris RH, Evans HJ (eds) Biological Nitrogen Fixation. Chapman and Hall, New York, pp 560–597

    Google Scholar 

  • Matsumura P, Rydel JJ, Linzmeier R, Vacante D (1984) Overexpression and sequence of the Escherichia coli cheY gene and biochemical activities of the CheY protein. J Bacteriol 160:36–41

    Google Scholar 

  • Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Mutoh N, Simon MI (1986) Nucleotide sequence corresponding to five chemotaxis genes in Escherichia coli. J Bacteriol 165:161–166

    Google Scholar 

  • Nagasawa S, Tokishita S, Aiba H, Mizuno T (1992) A novel sensor-regulator protein that belongs to the homologous family of signal-transduction proteins involved in adaptive responses in Escherichia coli. Mol Microbiol 6:799–807

    Google Scholar 

  • Ninfa AJ (1991) Protein phosphorylation and the regulation of cellular processes by the homologous two-component regulatory systems of bacteria. Genet Eng 13:39–72

    Google Scholar 

  • Ninfa AJ, Ninfa EG, Lupas AN, Stock A, Magasanik B, Stock J (1988) Crosstalk between bacterial chemotaxis signal transduction proteins and regulators of transcription of the Ntr regulon evidence that nitrogen assimilation and chemotaxis are controlled by a common phosphotransfer mechanism. Proc Natl Acad USA 85:5492–5496

    Google Scholar 

  • Parkinson JS, Kofoid EC (1992) Communication modules in bacterial signaling proteins. Annu Rev Genet 26:71–112

    Google Scholar 

  • Price NPJ, Relic B, Talmont E, Lewin A, Promé D, Pueppke SG, Maillet F, Dénarié J, Promé JC, Broughton WJ (1992) Broad-host-range Rhizobium species strain NGR234 secretes a family of carbamoylated, and fucosylated, nodulation signals that are O-acetylated or sulphated. Mol Microbiol 6:3575–3584

    Google Scholar 

  • Rabin RS, Stewart V (1992) Either of two functionally redundant sensor proteins, NarX and NarQ, is sufficient for nitrate regulation in Escherichia coli K-12. Proc Natl Acad Sci USA 89:8419–8423

    Google Scholar 

  • Ramseier TM, Winteler HV, Hennecke H (1991) Discovery and sequence analysis of bacterial genes involved in the biogenesis of c-type cytochromes. J Biol Chem 266:7793–7803

    Google Scholar 

  • Regensburger B, Hennecke H (1983) RNA polymerase from Rhizobium japonicum. Arch Microbiol 135:103–109

    Google Scholar 

  • Rostas K, Kondorosi E, Horvath B, Simoncsits A, Kondorosi A (1986) Conservation of extended promoter regions of nodulation genes in Rhizobium. Proc Natl Acad Sci USA 83:1757–1761

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sanjuan J, Carlson RW, Spaink HP, Bhat UR, Barbour WM, Glushka J, Stacey G (1992) A 2-O-methyl-fucose moiety is present in the lipo-oligosaccharide nodulation signal of Bradyrhizobium japonicum. Proc Natl Acad Sci USA 89:8789–8793

    Google Scholar 

  • Schlaman HRM, Okker RJH, Lugtenberg BJ (1992) Regulation of nodulation gene expression by NodD in rhizobia. J Bacteriol 174:5177–5182

    Google Scholar 

  • Schultze M, Quiclet-Sire B, Kondorosi E, Virelizier H, Glushka JN, Endre G, Géro SD, Kondorosi A (1992) Rhizobium meliloti produces a family of sulfated lipo-oligosaccharides exhibiting different degrees of plant host specificity. Proc Natl Acad Sci USA 89:192–196

    Google Scholar 

  • Simon R, Priefer U, Pühler A (1983) Vector plasmids for in vivo and in vitro manipulation of gram-negative bacteria. In: Pühler A (ed) Molecular genetics of the bacteria-plant interaction. Springer Verlag, Heidelberg, pp 98–106

    Google Scholar 

  • Simon R, Quandt J, Klipp W (1989) New derivatives of transposon Tn5 suitable for mobilization of replicons, generation of operon fusions and induction of genes in Gram-negative bacteria. Gene 80:161–169

    Google Scholar 

  • Spaink HP (1992) Rhizobial lipo-oligosaccharides: answers and questions. Plant Mol Biol 20:977–986

    Google Scholar 

  • Spaink HP, Sheeley DM, van Brussel AAN, Glushka J, York WS, Tak T, Geiger O, Kennedy EP, Reinhold VN, Lugtenberg BJJ (1991) A novel highly unsaturated fatty acid moiety of lipo-oligosaccharide signals determines host specificity of Rhizobium. Nature 354:125–130

    Google Scholar 

  • Spaink HP, Aarts A, Stacey G, Bloemberg GV, Lugtenberg BJJ, Kennedy EP (1992) Detection and separation of Rhizobium and Bradyrhizobium Nod metabolites using thin-layer chromatography. Mol Plant-Microbe Interact 5:72–80

    Google Scholar 

  • Stock JB, Ninfa AJ, Stock AM (1989) Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev 53:450–490

    Google Scholar 

  • Trach KA, Chapman JW, Piggot PJ, Hoch JA (1985) Deduced product of the stage 0 sporulation gene spoOF shares homology with the SpoOA, OmpR, and SfrA proteins. Proc Natl Acad Sci USA 82:7260–7264

    Google Scholar 

  • Wanner BL (1992) Is cross regulation by phosphorylation of twocomponent response regulator proteins important in bacteria? J Bacteriol 174:2053–2058

    Google Scholar 

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Communicated by A. Kondorosi

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Grob, P., Michel, P., Hennecke, H. et al. A novel response-regulator is able to suppress the nodulation defect of a Bradyrhizobium japonicum nodW mutant. Molec. Gen. Genet. 241, 531–541 (1993). https://doi.org/10.1007/BF00279895

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