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PMID: 3062575 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The DNA-binding domain of the transcriptional activator protein NifA resides in its carboxy terminus, recognises the upstream activator sequences of nif promoters and can be separated from the positive control function of NifA.

Nucleic acids research ·Vol. 16 ·No. 24 ·1988-12-23 ·Pages 11469-88

Morett E, Cannon W, Buck M

Abstract

The positive control protein NifA activates transcription of nitrogen fixation promoters in Klebsiella pneumoniae. NifA is believed to bind to specific sites, the upstream activator sequences (UAS's), of the nif promoters which it activates. We have now shown by mutation of the carboxy terminus of NifA that this is the DNA-binding domain and that the DNA-binding and positive activator functions of NifA can be separated. Mutational analysis of the nifH UAS and in vivo methylation protection analysis of the interaction of NifA with the nifH promoter demonstrates that the UAS is recognised by the carboxy terminus of NifA. The UAS's of K. pneumoniae nif promoters are also required for activation by the Rhizobium meliloti NifA indicating that this activator also possesses DNA-binding activity.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics,isolation & purification,physiology Base Sequence DNA Mutational Analysis DNA Replication DNA-Binding Proteins/genetics,isolation & purification Gene Expression Regulation Genes, Bacterial Klebsiella pneumoniae/genetics Molecular Sequence Data Nitrogen Fixation/genetics Promoter Regions, Genetic Rhizobium/genetics Sulfuric Acid Esters Transcription Factors/genetics,isolation & purification,physiology Transcription, Genetic
Chemicals
Bacterial Proteins DNA-Binding Proteins Sulfuric Acid Esters Transcription Factors dimethyl sulfate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Morett E
AFRC Institute of Plant Science Research, University of Sussex, Brighton, UK.
Cannon W
Buck M
References (29)
29 references, click to expand
  1. 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
  2. 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
  3. 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
  4. The operator-binding domain of lambda repressor: structure and DNA recognition.
    Nature. 1982 Jul 29;298(5873):443-7 PMID: 7088190
  5. 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
  6. Structure of the repressor-operator complex of bacteriophage 434.
    Nature. 1987 Apr 30-May 6;326(6116):846-52 PMID: 3553959
  7. 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
  8. Structure of catabolite gene activator protein at 2.9 A resolution suggests binding to left-handed B-DNA.
    Nature. 1981 Apr 30;290(5809):744-9 PMID: 6261152
  9. 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
  10. Mapping and expression of a regulatory nitrogen fixation gene (fixD) of Rhizobium meliloti.
    EMBO J. 1985 Nov;4(11):2751-6 PMID: 16453640
  11. Structure of the cro repressor from bacteriophage lambda and its interaction with DNA.
    Nature. 1981 Apr 30;290(5809):754-8 PMID: 6452580
  12. Nitrogen fixation by Klebsiella pneumoniae is inhibited by certain multicopy hybrid nif plasmids.
    J Bacteriol. 1983 Jan;153(1):45-56 PMID: 6336738
  13. Deletion analysis of Rhizobium meliloti symbiotic promoters.
    EMBO J. 1985 Oct;4(10):2419-24 PMID: 15929217
  14. Transcription of glnA in E. coli is stimulated by activator bound to sites far from the promoter.
    Cell. 1986 Jun 20;45(6):785-92 PMID: 2871943
  15. 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
  16. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli.
    Proc Natl Acad Sci U S A. 1986 Aug;83(16):5909-13 PMID: 2874557
  17. Positional requirements for the function of nif-specific upstream activator sequences.
    Mol Gen Genet. 1987 Nov;210(1):140-4 PMID: 3323835
  18. Mutants of the catabolite activator protein of Escherichia coli that are specifically deficient in the gene-activation function.
    Proc Natl Acad Sci U S A. 1987 Dec;84(23):8315-9 PMID: 2825186
  19. Regulation of nitrogen fixation genes.
    Annu Rev Genet. 1986;20:567-91 PMID: 3545064
  20. Protein kinase and phosphoprotein phosphatase activities of nitrogen regulatory proteins NTRB and NTRC of enteric bacteria: roles of the conserved amino-terminal domain of NTRC.
    Proc Natl Acad Sci U S A. 1988 Jul;85(14):4976-80 PMID: 2839825
  21. Transcriptional activation of the Klebsiella pneumoniae nitrogenase promoter may involve DNA loop formation.
    Mol Microbiol. 1987 Sep;1(2):243-9 PMID: 2835583
  22. 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
  23. Expression and functional analysis of the Rhizobium meliloti nifA gene.
    EMBO J. 1988 Jan;7(1):7-14 PMID: 16453824
  24. 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
  25. The three-dimensional structure of trp repressor.
    Nature. 1985 Oct 31-Nov 6;317(6040):782-6 PMID: 3903514
  26. 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
  27. Tandem promoters determine regulation of the Klebsiella pneumoniae glutamine synthetase (glnA) gene.
    Nucleic Acids Res. 1984 Oct 25;12(20):7811-30 PMID: 6149519
  28. Transcription of a Rhizobium leguminosarum biovar phaseoli gene needed for melanin synthesis is activated by nifA of Rhizobium and Klebsiella pneumoniae.
    Mol Microbiol. 1988 May;2(3):331-7 PMID: 3041240
  29. 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
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/16.24.11469
Published
1988-12-23
Pages
11469-88
Language
English
Region
England
NLM ID
0411011
PMCID
PMC339059
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/16.24.11469 PMC Full Text View journal details
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