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PMID: 2849102 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

NifA-dependent in vivo protection demonstrates that the upstream activator sequence of nif promoters is a protein binding site.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 24 ·1988-12-00 ·Pages 9401-5

Morett E, Buck M

Abstract

Primer-extension analysis of the Klebsiella pneumoniae nifH promoter was used to determine changes in the accessibility of the promoter DNA to methylation after exposure of growing cells to dimethyl sulfate. Four guanine residues present in the nifH upstream activator sequence (UAS), the proposed NifA binding site, were protected from methylation and two guanine residues were hypermethylated when the transcriptional activator protein NifA was present in the cells. The interaction detected at the nifH UAS was independent of the alternative sigma factor NtrA required for transcription of the nifH and other nif promoters. Mutations within the nifH UAS that diminish NifA-dependent transcriptional activation reduced the interaction at the UAS. It seems likely that the pattern of methylation protection observed in the nifH UAS is the result of NifA binding.

MeSH Terms
Bacterial Proteins/analysis,genetics Base Sequence Binding Sites DNA, Bacterial/analysis Klebsiella pneumoniae/genetics Methylation Molecular Sequence Data Mutation Plasmids Promoter Regions, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Morett E
Agricultural & Food Research Council, University of Sussex, Brighton, United Kingdom.
Buck M
References (30)
30 references, click to expand
  1. Overproduction of phage lambda repressor under control of the lac promotor of Escherichia coli.
    Mol Gen Genet. 1976 Nov 17;148(3):243-50 PMID: 796661
  2. The role of activator binding sites in transcriptional control of the divergently transcribed nifF and nifLA promoters from Klebsiella pneumoniae.
    Mol Microbiol. 1988 Jul;2(4):433-42 PMID: 3139967
  3. DNA-binding site of lac repressor probed by dimethylsulfate methylation of lac operator.
    J Mol Biol. 1979 Aug 25;132(4):709-28 PMID: 231106
  4. The products of glnL and glnG are bifunctional regulatory proteins.
    Mol Gen Genet. 1982;188(2):325-33 PMID: 6130464
  5. An open reading frame upstream from the nifH gene of Klebsiella pneumoniae.
    Nucleic Acids Res. 1983 Jun 25;11(12):4241-50 PMID: 6306580
  6. Mutations affecting regulation of the Klebsiella pneumoniae nifH (nitrogenase reductase) promotor.
    J Bacteriol. 1984 Jan;157(1):143-7 PMID: 6360993
  7. Nitrogen fixation specific regulatory genes of Klebsiella pneumoniae and Rhizobium meliloti share homology with the general nitrogen regulatory gene ntrC of K. pneumoniae.
    Nucleic Acids Res. 1985 Jun 25;13(12):4539-55 PMID: 2989799
  8. Nitrogen regulation in Salmonella typhimurium. Identification of an ntrC protein-binding site and definition of a consensus binding sequence.
    EMBO J. 1985 Feb;4(2):539-47 PMID: 2862031
  9. Site-directed mutagenesis of the Klebsiella pneumoniae nifL and nifH promoters and in vivo analysis of promoter activity.
    Nucleic Acids Res. 1985 Nov 11;13(21):7621-38 PMID: 3906564
  10. Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a sigma factor.
    Proc Natl Acad Sci U S A. 1985 Nov;82(22):7525-9 PMID: 2999766
  11. Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8453-7 PMID: 2867543
  12. Interaction of purified NtrC protein with nitrogen regulated promoters from Klebsiella pneumoniae.
    Mol Gen Genet. 1985;201(3):492-8 PMID: 3911027
  13. Deletion analysis of the Klebsiella pneumoniae nitrogenase promoter: importance of spacing between conserved sequences around positions -12 and -24 for activation by the nifA and ntrC (glnG) products.
    J Bacteriol. 1986 May;166(2):545-51 PMID: 3009407
  14. Sequence and domain relationships of ntrC and nifA from Klebsiella pneumoniae: homologies to other regulatory proteins.
    EMBO J. 1986 Feb;5(2):441-7 PMID: 3011408
  15. High-resolution analysis of lac transcription complexes inside cells.
    Biochemistry. 1986 Sep 9;25(18):5051-7 PMID: 3533143
  16. Regulation of nitrogen fixation genes.
    Annu Rev Genet. 1986;20:567-91 PMID: 3545064
  17. Organization and partial sequence of a DNA region of the Rhizobium leguminosarum symbiotic plasmid pRL6JI containing the genes fixABC, nifA, nifB and a novel open reading frame.
    Nucleic Acids Res. 1987 Jan 12;15(1):31-49 PMID: 3029674
  18. In vitro transcription of the nitrogen fixation regulatory operon nifLA of Klebsiella pneumoniae.
    J Bacteriol. 1987 Jun;169(6):2876-80 PMID: 3294810
  19. Initiation of transcription at the bacterial glnAp2 promoter by purified E. coli components is facilitated by enhancers.
    Cell. 1987 Sep 25;50(7):1039-46 PMID: 3304660
  20. Mutations that create new promoters suppress the sigma 54 dependence of glnA transcription in Escherichia coli.
    J Bacteriol. 1987 Sep;169(9):4279-84 PMID: 2887548
  21. Deduced products of C4-dicarboxylate transport regulatory genes of Rhizobium leguminosarum are homologous to nitrogen regulatory gene products.
    Nucleic Acids Res. 1987 Oct 12;15(19):7921-34 PMID: 3671068
  22. Mutational analysis of upstream sequences required for transcriptional activation of the Klebsiella pneumoniae nifH promoter.
    Nucleic Acids Res. 1987 Dec 10;15(23):9945-56 PMID: 3320958
  23. Involvement of the ntrA gene product in the anaerobic metabolism of Escherichia coli.
    Mol Gen Genet. 1987 Dec;210(3):535-42 PMID: 3323848
  24. Essential and non-essential domains in the Bradyrhizobium japonicum NifA protein: identification of indispensable cysteine residues potentially involved in redox reactivity and/or metal binding.
    Nucleic Acids Res. 1988 Mar 25;16(5):2207-24 PMID: 3357773
  25. Transcriptional activation of the Klebsiella pneumoniae nitrogenase promoter may involve DNA loop formation.
    Mol Microbiol. 1987 Sep;1(2):243-9 PMID: 2835583
  26. The effect on the function of the transcriptional activator NtrC from Klebsiella pneumoniae of mutations in the DNA-recognition helix.
    Nucleic Acids Res. 1988 May 11;16(9):4025-39 PMID: 3287338
  27. Requirements for transcriptional activation in vitro of the nitrogen-regulated glnA and nifLA promoters from Klebsiella pneumoniae: dependence on activator concentration.
    Mol Microbiol. 1987 Jul;1(1):92-100 PMID: 3330758
  28. Nucleotide sequence and mutagenesis of the nifA gene from Azotobacter vinelandii.
    Mol Microbiol. 1988 May;2(3):315-21 PMID: 2840552
  29. Over-production and characterization of the nifA gene product of Klebsiella pneumoniae--the transcriptional activator of nif gene expression.
    J Gen Microbiol. 1988 Feb;134(2):425-32 PMID: 2844961
  30. Contacts between Escherichia coli RNA polymerase and a lac operon promoter.
    Proc Natl Acad Sci U S A. 1978 Nov;75(11):5314-8 PMID: 364474
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.24.9401
Published
1988-12-00
Pages
9401-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC282760
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.85.24.9401 PMC Full Text View journal details
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