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. 1982 Apr;150(1):293–297. doi: 10.1128/jb.150.1.293-297.1982

Intragenic complementation by the nifJ-coded protein of Klebsiella pneumoniae.

G Stacey, J Zhu, V K Shah, S C Shen, W J Brill
PMCID: PMC220112  PMID: 7037747

Abstract

A single mutation, nifC1005 (Jin et al. Sci. Sin. 23:108-118, 1980), located between nifH and nifJ in the nif cluster of Klebsiella pneumoniae, genetically complemented mutations in each of the 17 known nif genes. This suggested that the mutation is located in a new nif gene. We showed by complementation analyses that only 3 of 12 nifJ mutations tested were complemented by nifC1005. Nitrogenase activity in cell extracts of the mutant with nifC1005 as well as NifJ- mutants was stimulated by the addition of the iron-molybdenum cofactor or nitrogenase component I. The molecular weight of the native NifJ protein is approximately 257,000--a dimer of identical subunits. Some nifC-/nifJ- or nifJ-/nifJ- merodiploids produced active but unstable nifJ proteins. Fine-structure mapping placed the nifC1005 allele within the nifJ gene bounded on both sides by well-characterized nifJ mutations. This indicates that the nifC1005 does not define a separate gene from nifJ. The data are consistent with the occurrence of intragenic complementation between two defective nifJ polypeptides. This explains the isolated examples of genetic complementation between the nifC1005 mutation and certain nifJ mutations.

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Selected References

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

  1. Bachhuber M., Brill W. J., Howe M. M. Use of bacteriophage Mu to isolate deletions in the his-nif region of Klebsiella pneumoniae. J Bacteriol. 1976 Dec;128(3):749–753. doi: 10.1128/jb.128.3.749-753.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brill W. J. Biochemical genetics of nitrogen fixation. Microbiol Rev. 1980 Sep;44(3):449–467. doi: 10.1128/mr.44.3.449-467.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CRICK F. H., ORGEL L. E. THE THEORY OF INTER-ALLELIC COMPLEMENTATION. J Mol Biol. 1964 Jan;8:161–165. doi: 10.1016/s0022-2836(64)80156-x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Hill S., Kavanagh E. P. Roles of nifF and nifJ gene products in electron transport to nitrogenase in Klebsiella pneumoniae. J Bacteriol. 1980 Feb;141(2):470–475. doi: 10.1128/jb.141.2.470-475.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. MacNeil T., Brill W. J., Howe M. M. Bacteriophage mu-induced deletions in a plasmid containing the nif (N2 fixation) genes of Klebsiella pneumoniae. J Bacteriol. 1978 Jun;134(3):821–829. doi: 10.1128/jb.134.3.821-829.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Nagatani H. H., Shah V. K., Brill W. J. Activation of inactive nitrogenase by acid-treated component I. J Bacteriol. 1974 Nov;120(2):697–701. doi: 10.1128/jb.120.2.697-701.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nieva-Gómez D., Roberts G. P., Klevickis S., Brill W. J. Electron transport to nitrogenase in Klebsiella pneumoniae. Proc Natl Acad Sci U S A. 1980 May;77(5):2555–2558. doi: 10.1073/pnas.77.5.2555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pienkos P. T., Brill W. J. Molybdenum accumulation and storage in Klebsiella pneumoniae and Azotobacter vinelandii. J Bacteriol. 1981 Feb;145(2):743–751. doi: 10.1128/jb.145.2.743-751.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Runzhi J., Yide H., Meijing S., Biao C. J., Dean G., Bingfu S., Wujun L., Shanjiong S. Complementation analysis and characterization of the nitrogen fixation genes, nifH, nifC and nifJ in Klebsiella pneumoniae. Sci Sin. 1980 Jan;23(1):108–118. [PubMed] [Google Scholar]
  17. Shah V. K., Brill W. J. Isolation of an iron-molybdenum cofactor from nitrogenase. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3249–3253. doi: 10.1073/pnas.74.8.3249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shah V. K., Brill W. J. Nitrogenase. IV. Simple method of purification to homogeneity of nitrogenase components from Azotobacter vinelandii. Biochim Biophys Acta. 1973 May 30;305(2):445–454. doi: 10.1016/0005-2728(73)90190-4. [DOI] [PubMed] [Google Scholar]
  19. Shah V. K., Davis L. C., Brill W. J. Nitrogenase. I. Repression and derepression of the iron-molybdenum and iron proteins of nitrogenase in Azotobacter vinelandii. Biochim Biophys Acta. 1972 Feb 28;256(2):498–511. doi: 10.1016/0005-2728(72)90078-3. [DOI] [PubMed] [Google Scholar]
  20. St John R. T., Johnston H. M., Seidman C., Garfinkel D., Gordon J. K., Shah V. K., Brill W. J. Biochemistry and genetics of Klebsiella pneumoniae mutant strains unable to fix N2. J Bacteriol. 1975 Mar;121(3):759–765. doi: 10.1128/jb.121.3.759-765.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ullmann A., Perrin D., Jacob F., Monod J. Identification par complémentation in vitro et purification d'un segment peptidique de la beta-galatosidase d'escherichia coli. J Mol Biol. 1965 Jul;12(3):918–923. doi: 10.1016/s0022-2836(65)80338-2. [DOI] [PubMed] [Google Scholar]

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