Abstract
In our studies of nif gene regulation, we have observed that certain hybrid nif plasmids drastically inhibit the expression of the chromosomal nif genes of Klebsiella pneumonia. Wild-type (Nif+) K. pneumoniae strains that acquire certain hybrid nif plasmids also acquire the Nif- phenotype; these strains lose 90 to 99% of all detectable nitrogen fixation activity and grow poorly (or not at all) on solid media with N2 as the sole nitrogen source. We describe experiments which defined this inhibition of the Nif+ phenotype by hybrid nif plasmids and identify and characterize four nif DNA regions associated with this inhibition. We show that plasmids carrying these nif regions could recombine with, but not complement, nif chromosomal mutations. Our results suggest that inhibition of the Nif+ phenotype will provide a useful bioassay for some of the factors that mediate nif gene expression.
Full text
PDF











Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Appelbaum E. R., Kramer R. A. Restriction mapping of deletions in the nif region of the Klebsiella pneumoniae chromosome. Mol Gen Genet. 1980;179(2):349–354. doi: 10.1007/BF00425463. [DOI] [PubMed] [Google Scholar]
- Ausubel F. M., Bird S. C., Durbin K. J., Janssen K. A., Margolskee R. F., Peskin A. P. Glutamine synthetase mutations which affect expression of nitrogen fixation genes in Klebsiella pneumoniae. J Bacteriol. 1979 Nov;140(2):597–606. doi: 10.1128/jb.140.2.597-606.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Backman K., Ptashne M., Gilbert W. Construction of plasmids carrying the cI gene of bacteriophage lambda. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4174–4178. doi: 10.1073/pnas.73.11.4174. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
- Cannon F. C., Riedel G. E., Ausubel F. M. Overlapping sequences of Klebsiella pneumoniae nifDNA cloned and characterized. Mol Gen Genet. 1979 Jul 2;174(1):59–66. doi: 10.1007/BF00433306. [DOI] [PubMed] [Google Scholar]
- Cannon F. C., Riedel G. E., Ausubel F. M. Recombinant plasmid that carries part of the nitrogen fixation (nif) gene cluster of Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2963–2967. doi: 10.1073/pnas.74.7.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dixon R., Eady R. R., Espin G., Hill S., Iaccarino M., Kahn D., Merrick M. Analysis of regulation of Klebsiella pneumoniae nitrogen fixation (nif) gene cluster with gene fusions. Nature. 1980 Jul 10;286(5769):128–132. doi: 10.1038/286128a0. [DOI] [PubMed] [Google Scholar]
- Dixon R., Kennedy C., Kondorosi A., Krishnapillai V., Merrick M. Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol Gen Genet. 1977 Nov 29;157(2):189–198. doi: 10.1007/BF00267397. [DOI] [PubMed] [Google Scholar]
- Dugaiczyk A., Boyer H. W., Goodman H. M. Ligation of EcoRI endonuclease-generated DNA fragments into linear and circular structures. J Mol Biol. 1975 Jul 25;96(1):171–184. doi: 10.1016/0022-2836(75)90189-8. [DOI] [PubMed] [Google Scholar]
- Elmerich C., Houmard J., Sibold L., Manheimer I., Charpin N. Genetic and biochemical analysis of mutants induced by bacteriophage Mu DNA integration into Klebsiella pneumoniae nitrogen fixation genes. Mol Gen Genet. 1978 Oct 4;165(2):181–189. doi: 10.1007/BF00269905. [DOI] [PubMed] [Google Scholar]
- Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heyneker H. L., Shine J., Goodman H. M., Boyer H. W., Rosenberg J., Dickerson R. E., Narang S. A., Itakura K., Lin S., Riggs A. D. Synthetic lac operator DNA is functional in vivo. Nature. 1976 Oct 28;263(5580):748–752. doi: 10.1038/263748a0. [DOI] [PubMed] [Google Scholar]
- Hill S., Kennedy C., Kavanagh E., Goldberg R. B., Hanau R. Nitrogen fixation gene (nifL) involved in oxygen regulation of nitrogenase synthesis in K. pneumoniae. Nature. 1981 Apr 2;290(5805):424–426. doi: 10.1038/290424a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Klein R. D., Selsing E., Wells R. D. A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid. 1980 Jan;3(1):88–91. doi: 10.1016/s0147-619x(80)90037-2. [DOI] [PubMed] [Google Scholar]
- MAHL M. C., WILSON P. W., FIFE M. A., EWING W. H. NITROGEN FIXATION BY MEMBERS OF THE TRIBE KLEBSIELLEAE. J Bacteriol. 1965 Jun;89:1482–1487. doi: 10.1128/jb.89.6.1482-1487.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacNeil D., Brill W. J. Mutations in nif genes that cause Klebsiella pneumoniae to be derepressed for nitrogenase synthesis in the presence of ammonium. J Bacteriol. 1980 Nov;144(2):744–751. doi: 10.1128/jb.144.2.744-751.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacNeil D., Howe M. M., Brill W. J. Isolation and characterization of lambda specialized transducing bacteriophages carrying Klebsiella pneumoniae nif genes. J Bacteriol. 1980 Mar;141(3):1264–1271. doi: 10.1128/jb.141.3.1264-1271.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacNeil D., Zhu J., Brill W. J. Regulation of nitrogen fixation in Klebsiella pneumoniae: isolation and characterization of strains with nif-lac fusions. J Bacteriol. 1981 Jan;145(1):348–357. doi: 10.1128/jb.145.1.348-357.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacNeil T., MacNeil D., Roberts G. P., Supiano M. A., Brill W. J. Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae. J Bacteriol. 1978 Oct;136(1):253–266. doi: 10.1128/jb.136.1.253-266.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
- Marians K. J., Wu R., Stawinski J., Hozumi T., Narang S. A. Cloned synthetic lac operator DNA is biologically active. Nature. 1976 Oct 28;263(5580):744–748. doi: 10.1038/263744a0. [DOI] [PubMed] [Google Scholar]
- Merrick M., Filser M., Dixon R., Elmerich C., Sibold L., Houmard J. The use of translocatable genetic elements to construct a fine-structure map of the Klebsiella pneumoniae nitrogen fixation (nif) gene cluster. J Gen Microbiol. 1980 Apr;117(2):509–520. doi: 10.1099/00221287-117-2-509. [DOI] [PubMed] [Google Scholar]
- Merrick M., Filser M., Kennedy C., Dixon R. Polarity of mutations induced by insertion of transposons Tn5, Tn7 and Tn10 into the nif gene cluster of Klebsiella pneumoniae. Mol Gen Genet. 1978 Sep 20;165(1):103–111. doi: 10.1007/BF00270382. [DOI] [PubMed] [Google Scholar]
- Riedel G. E., Ausubel F. M., Cannon F. C. Physical map of chromosomal nitrogen fixation (nif) genes of Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2866–2870. doi: 10.1073/pnas.76.6.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roberts G. P., Brill W. J. Gene-product relationships of the nif regulon of Klebsiella pneumoniae. J Bacteriol. 1980 Oct;144(1):210–216. doi: 10.1128/jb.144.1.210-216.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roberts G. P., MacNeil T., MacNeil D., Brill W. J. Regulation and characterization of protein products coded by the nif (nitrogen fixation) genes of Klebsiella pneumoniae. J Bacteriol. 1978 Oct;136(1):267–279. doi: 10.1128/jb.136.1.267-279.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sakano H., Rogers J. H., Hüppi K., Brack C., Traunecker A., Maki R., Wall R., Tonegawa S. Domains and the hinge region of an immunoglobulin heavy chain are encoded in separate DNA segments. Nature. 1979 Feb 22;277(5698):627–633. doi: 10.1038/277627a0. [DOI] [PubMed] [Google Scholar]
- Scott K. F., Rolfe B. G., Shine J. Biological nitrogen fixation: primary structure of the Klebsiella pneumoniae nifH and nifD genes. J Mol Appl Genet. 1981;1(1):71–81. [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Streicher S. L., Shanmugam K. T., Ausubel F., Morandi C., Goldberg R. B. Regulation of nitrogen fixation in Klebsiella pneumoniae: evidence for a role of glutamine synthetase as a regulator of nitrogenase synthesis. J Bacteriol. 1974 Nov;120(2):815–821. doi: 10.1128/jb.120.2.815-821.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tyler B. Regulation of the assimilation of nitrogen compounds. Annu Rev Biochem. 1978;47:1127–1162. doi: 10.1146/annurev.bi.47.070178.005403. [DOI] [PubMed] [Google Scholar]
- Zhu J., Brill W. J. Temperature sensitivity of the regulation of nitrogenase synthesis by Klebsiella pneumoniae. J Bacteriol. 1981 Feb;145(2):1116–1118. doi: 10.1128/jb.145.2.1116-1118.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]