Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1989 Dec;55(12):3051–3057. doi: 10.1128/aem.55.12.3051-3057.1989

Identification of a Locus Upstream from the Hydrogenase Structural Genes That Is Involved in Hydrogenase Expression in Bradyrhizobium japonicum

Patricia D Novak 1, Robert J Maier 1,*
PMCID: PMC203222  PMID: 16348066

Abstract

A locus involved in the expression of the uptake hydrogenase system of Bradyrhizobium japonicum was identified adjacent to genes encoding the hydrogenase subunits. A cloned fragment of DNA was used to complement to autotrophy a Hup putative regulatory mutant of B. japonicum. The mutant strain lacked hydrogenase activity and synthesized low levels of the large subunit of hydrogenase as determined by Western gels. Tn5-induced mutagenesis located the region within the fragment which was necessary for complementation of the mutant phenotype. The locus identified is adjacent to that encoding the small subunit of hydrogenase; its right border is <0.5 kilobase upstream from the hydrogenase transcriptional start site, and its left border is between 1 and 2.5 kilobases from that start site. However, the locus is outside the region previously shown to contain hup-related genes of B. japonicum. Thus, the localization of this gene describes a previously unidentified hup-related gene on a region of DNA not previously shown to contain hup-specific DNA.

Full text

PDF
3051

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agarwal A. K., Keister D. L. Physiology of Ex Planta Nitrogenase Activity in Rhizobium japonicum. Appl Environ Microbiol. 1983 May;45(5):1592–1601. doi: 10.1128/aem.45.5.1592-1601.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bishop P. E., Guevara J. G., Engelke J. A., Evans H. J. Relation between Glutamine Synthetase and Nitrogenase Activities in the Symbiotic Association between Rhizobium japonicum and Glycine max. Plant Physiol. 1976 Apr;57(4):542–546. doi: 10.1104/pp.57.4.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  4. Cantrell M. A., Haugland R. A., Evans H. J. Construction of a Rhizobium japonicum gene bank and use in isolation of a hydrogen uptake gene. Proc Natl Acad Sci U S A. 1983 Jan;80(1):181–185. doi: 10.1073/pnas.80.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X. W., Finlay D. R., Guiney D., Helinski D. R. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. doi: 10.1016/0147-619x(85)90068-x. [DOI] [PubMed] [Google Scholar]
  6. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. doi: 10.1016/0003-2697(81)90296-7. [DOI] [PubMed] [Google Scholar]
  8. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
  10. Hanus F. J., Maier R. J., Evans H. J. Autotrophic growth of H2-uptake-positive strains of Rhizobium japonicum in an atmosphere supplied with hydrogen gas. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1788–1792. doi: 10.1073/pnas.76.4.1788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Haugland R. A., Cantrell M. A., Beaty J. S., Hanus F. J., Russell S. A., Evans H. J. Characterization of Rhizobium japonicum hydrogen uptake genes. J Bacteriol. 1984 Sep;159(3):1006–1012. doi: 10.1128/jb.159.3.1006-1012.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hom S. S., Graham L. A., Maier R. J. Isolation of genes (nif/hup cosmids) involved in hydrogenase and nitrogenase activities in Rhizobium japonicum. J Bacteriol. 1985 Mar;161(3):882–887. doi: 10.1128/jb.161.3.882-887.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hom S. S., Novak P. D., Maier R. J. Transposon Tn5-Generated Bradyrhizobium japonicum Mutants Unable To Grow Chemoautotrophically with H(2). Appl Environ Microbiol. 1988 Feb;54(2):358–363. doi: 10.1128/aem.54.2.358-363.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jorgensen R. A., Rothstein S. J., Reznikoff W. S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. doi: 10.1007/BF00267254. [DOI] [PubMed] [Google Scholar]
  16. Lambert G. R., Cantrell M. A., Hanus F. J., Russell S. A., Haddad K. R., Evans H. J. Intra- and interspecies transfer and expression of Rhizobium japonicum hydrogen uptake genes and autotrophic growth capability. Proc Natl Acad Sci U S A. 1985 May;82(10):3232–3236. doi: 10.1073/pnas.82.10.3232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lambert G. R., Harker A. R., Cantrell M. A., Hanus F. J., Russell S. A., Haugland R. A., Evans H. J. Symbiotic Expression of Cosmid-Borne Bradyrhizobium japonicum Hydrogenase Genes. Appl Environ Microbiol. 1987 Feb;53(2):422–428. doi: 10.1128/aem.53.2.422-428.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lepo J. E., Hanus F. J., Evans H. J. Chemoautotrophic growth of hydrogen-uptake-positive strains of Rhizobium japonicum. J Bacteriol. 1980 Feb;141(2):664–670. doi: 10.1128/jb.141.2.664-670.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lepo J. E., Hickok R. E., Cantrell M. A., Russell S. A., Evans H. J. Revertible hydrogen uptake-deficient mutants of Rhizobium japonicum. J Bacteriol. 1981 May;146(2):614–620. doi: 10.1128/jb.146.2.614-620.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lim S. T. Determination of Hydrogenase in Free-living Cultures of Rhizobium japonicum and Energy Efficiency of Soybean Nodules. Plant Physiol. 1978 Oct;62(4):609–611. doi: 10.1104/pp.62.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maier R. J., Merberg D. M. Rhizobium japonicum mutants that are hypersensitive to repression of H2 uptake by oxygen. J Bacteriol. 1982 Apr;150(1):161–167. doi: 10.1128/jb.150.1.161-167.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maier R. J. Rhizobium japonicum mutant strains unable to grow chemoautotrophically with H2. J Bacteriol. 1981 Jan;145(1):533–540. doi: 10.1128/jb.145.1.533-540.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moshiri F., Maier R. J. Conformational changes in the membrane-bound hydrogenase of Bradyrhizobium japonicum. Evidence that the redox state of the enzyme affects its accessibility to protease and membrane-impermeant reagents. J Biol Chem. 1988 Nov 25;263(33):17809–17816. [PubMed] [Google Scholar]
  24. Moshiri F., Stults L., Novak P., Maier R. J. Nif- Hup- mutants of Rhizobium japonicum. J Bacteriol. 1983 Aug;155(2):926–929. doi: 10.1128/jb.155.2.926-929.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Brian M. R., Maier R. J. Electron transport components involved in hydrogen oxidation in free-living Rhizobium japonicum. J Bacteriol. 1982 Oct;152(1):422–430. doi: 10.1128/jb.152.1.422-430.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Polacheck I., Cabib E. A simple procedure for protein determination by the Lowry method in dilute solutions and in the presence of interfering substances. Anal Biochem. 1981 Nov 1;117(2):311–314. doi: 10.1016/0003-2697(81)90784-3. [DOI] [PubMed] [Google Scholar]
  27. Reeve J. N., Beckler G. S., Cram D. S., Hamilton P. T., Brown J. W., Krzycki J. A., Kolodziej A. F., Alex L., Orme-Johnson W. H., Walsh C. T. A hydrogenase-linked gene in Methanobacterium thermoautotrophicum strain delta H encodes a polyferredoxin. Proc Natl Acad Sci U S A. 1989 May;86(9):3031–3035. doi: 10.1073/pnas.86.9.3031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
  29. Sayavedra-Soto L. A., Powell G. K., Evans H. J., Morris R. O. Nucleotide sequence of the genetic loci encoding subunits of Bradyrhizobium japonicum uptake hydrogenase. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8395–8399. doi: 10.1073/pnas.85.22.8395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stults L. W., Moshiri F., Maier R. J. Aerobic purification of hydrogenase from Rhizobium japonicum by affinity chromatography. J Bacteriol. 1986 Jun;166(3):795–800. doi: 10.1128/jb.166.3.795-800.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stults L. W., O'Hara E. B., Maier R. J. Nickel is a component of hydrogenase in Rhizobium japonicum. J Bacteriol. 1984 Jul;159(1):153–158. doi: 10.1128/jb.159.1.153-158.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Winberg G., Hammarskjöld M. L. Isolation of DNA from agarose gels using DEAE-paper. Application to restriction site mapping of adenovirus type 16 DNA. Nucleic Acids Res. 1980 Jan 25;8(2):253–264. doi: 10.1093/nar/8.2.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zuber M., Harker A. R., Sultana M. A., Evans H. J. Cloning and expression of Bradyrhizobium japonicum uptake hydrogenase structural genes in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7668–7672. doi: 10.1073/pnas.83.20.7668. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES