Home LiteratureArticle Details
PMID: 8981978 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of fhlA mutations resulting in ligand-independent transcriptional activation and ATP hydrolysis.

Journal of bacteriology ·Vol. 179 ·No. 1 ·1997-01-00 ·Pages 41-5

Korsa I, Böck A

Abstract

The FhlA protein belongs to the NtrC family of transcriptional regulators. It induces transcription from the -12/-24 promoters of the genes of the formate regulon by sigma54 RNA polymerase. FhlA is activated by binding of the ligand formate and does not require phosphorylation. A mutational analysis of the fhLA gene portion coding for the A and C domains was conducted with the aim of gaining information on the interaction between formate binding and ATP hydrolysis plus transcription activation. Four mutations were identified, all located in the A domain; one of them rendered transcription completely independent from the presence of formate, and the others conferred a semiconstitutive phenotype. The FhlA protein of one of the semiconstitutive variants was purified. Catalytic efficiency of ATP hydrolysis of the mutant FhlA was increased in the absence of formate in the same manner as formate influences the activity of wild-type FhlA. Moreover, in vitro transcription occurred at much lower threshold concentrations of the mutant protein and of nucleoside triphosphates than with the wild-type FhlA.

MeSH Terms
Adenosine Triphosphate/metabolism Escherichia coli/genetics,metabolism Escherichia coli Proteins Formates/pharmacology Gene Expression Regulation, Bacterial/drug effects Genes, Bacterial/genetics Hydrolysis Kinetics Ligands Mutation Trans-Activators/genetics,isolation & purification,metabolism Transcriptional Activation/drug effects
Chemicals
Escherichia coli Proteins Formates Ligands Trans-Activators formic acid fhlA protein, E coli Adenosine Triphosphate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Korsa I
Lehrstuhl für Mikrobiologie der Universität München, Munich, Germany.
Böck A
References (39)
39 references, click to expand
  1. Analysis of the hydA locus of Escherichia coli: two genes (hydN and hypF) involved in formate and hydrogen metabolism.
    Arch Microbiol. 1996 May;165(5):333-41 PMID: 8661925
  2. Mutational analysis of the operon (hyc) determining hydrogenase 3 formation in Escherichia coli.
    Mol Microbiol. 1992 Jun;6(11):1523-32 PMID: 1625581
  3. Cloning and nucleotide sequence of the gene encoding the positive regulator (DmpR) of the phenol catabolic pathway encoded by pVI150 and identification of DmpR as a member of the NtrC family of transcriptional activators.
    J Bacteriol. 1993 Mar;175(6):1596-604 PMID: 8449869
  4. Sensing of aromatic compounds by the DmpR transcriptional activator of phenol-catabolizing Pseudomonas sp. strain CF600.
    J Bacteriol. 1994 Mar;176(6):1555-60 PMID: 8132448
  5. Genetic evidence for activation of the positive transcriptional regulator Xy1R, a member of the NtrC family of regulators, by effector binding.
    J Biol Chem. 1994 Mar 18;269(11):8059-62 PMID: 8132529
  6. Purification and DNA-binding properties of FHLA, the transcriptional activator of the formate hydrogenlyase system from Escherichia coli.
    J Biol Chem. 1994 Jul 29;269(30):19590-6 PMID: 8034727
  7. Regulated expression in vitro of genes coding for formate hydrogenlyase components of Escherichia coli.
    J Biol Chem. 1994 Jul 29;269(30):19597-604 PMID: 8034728
  8. The major dimerization determinants of the nitrogen regulatory protein NTRC from enteric bacteria lie in its carboxy-terminal domain.
    J Mol Biol. 1994 Aug 12;241(2):233-45 PMID: 8057363
  9. Characterisation of a protease from Escherichia coli involved in hydrogenase maturation.
    Eur J Biochem. 1995 Jan 15;227(1-2):545-50 PMID: 7851435
  10. Effector-mediated stimulation of ATPase activity by the sigma 54-dependent transcriptional activator FHLA from Escherichia coli.
    J Bacteriol. 1995 May;177(10):2798-803 PMID: 7751289
  11. Constitutive forms of the enhancer-binding protein NtrC: evidence that essential oligomerization determinants lie in the central activation domain.
    J Mol Biol. 1995 Jun 16;249(4):700-13 PMID: 7602583
  12. A common switch in activation of the response regulators NtrC and PhoB: phosphorylation induces dimerization of the receiver modules.
    EMBO J. 1995 Aug 1;14(15):3696-705 PMID: 7641688
  13. The bacterial enhancer-binding protein NTRC is a molecular machine: ATP hydrolysis is coupled to transcriptional activation.
    Genes Dev. 1995 Aug 15;9(16):2042-52 PMID: 7649482
  14. Activation of the transcriptional regulator XylR of Pseudomonas putida by release of repression between functional domains.
    Mol Microbiol. 1995 Apr;16(2):205-13 PMID: 7565083
  15. New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressureized atmosphere.
    Appl Environ Microbiol. 1976 Dec;32(6):781-91 PMID: 827241
  16. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  17. Analysis of regulation of Klebsiella pneumoniae nitrogen fixation (nif) gene cluster with gene fusions.
    Nature. 1980 Jul 10;286(5769):128-32 PMID: 6995849
  18. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074-8 PMID: 3156376
  19. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  20. The seleno-polypeptide of formic dehydrogenase (formate hydrogen-lyase linked) from Escherichia coli: genetic analysis.
    Arch Microbiol. 1985 May;141(4):359-63 PMID: 3160320
  21. 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
  22. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Methods Enzymol. 1987;154:367-82 PMID: 3323813
  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. Transformation of bacteria with plasmid DNA by electroporation.
    Anal Biochem. 1988 Apr;170(1):38-44 PMID: 3133958
  25. Nucleotide sequence of the regulatory gene xylR of the TOL plasmid from Pseudomonas putida.
    Gene. 1988 Jun 30;66(2):301-6 PMID: 3169574
  26. Characterization of a cis regulatory DNA element necessary for formate induction of the formate dehydrogenase gene (fdhF) of Escherichia coli.
    Mol Microbiol. 1989 Feb;3(2):187-95 PMID: 2668685
  27. Mutations in trans which affect the anaerobic expression of a formate dehydrogenase (fdhF) structural gene.
    Arch Microbiol. 1989;152(1):83-9 PMID: 2669674
  28. Characterization of divergent NtrA-dependent promoters in the anaerobically expressed gene cluster coding for hydrogenase 3 components of Escherichia coli.
    Mol Microbiol. 1990 Jan;4(1):13-20 PMID: 2181234
  29. Nucleotide sequence and expression of an operon in Escherichia coli coding for formate hydrogenlyase components.
    Mol Microbiol. 1990 Feb;4(2):231-43 PMID: 2187144
  30. Identification and sequence analysis of the gene encoding the transcriptional activator of the formate hydrogenlyase system of Escherichia coli.
    Mol Microbiol. 1990 Aug;4(8):1319-27 PMID: 2280686
  31. Molecular characterization of an operon (hyp) necessary for the activity of the three hydrogenase isoenzymes in Escherichia coli.
    Mol Microbiol. 1991 Jan;5(1):123-35 PMID: 1849603
  32. The central domain of Rhizobium leguminosarum DctD functions independently to activate transcription.
    J Bacteriol. 1992 Feb;174(4):1428-31 PMID: 1735730
  33. Mechanism of regulation of the formate-hydrogenlyase pathway by oxygen, nitrate, and pH: definition of the formate regulon.
    Mol Microbiol. 1991 Nov;5(11):2807-14 PMID: 1779767
  34. Phosphorylation of nitrogen regulator I of Escherichia coli induces strong cooperative binding to DNA essential for activation of transcription.
    Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5088-92 PMID: 1350679
  35. The prokaryotic enhancer binding protein NTRC has an ATPase activity which is phosphorylation and DNA dependent.
    EMBO J. 1992 Jun;11(6):2219-28 PMID: 1534752
  36. Direct regulation of the ATPase activity of the transcriptional activator DmpR by aromatic compounds.
    Mol Microbiol. 1995 Aug;17(3):505-13 PMID: 8559069
  37. The nucleotide concentration determines the specificity of in vitro transcription activation by the sigma 54-dependent activator FhlA.
    J Bacteriol. 1996 Jan;178(1):199-203 PMID: 8550417
  38. In vitro activities of an N-terminal truncated form of XylR, a sigma 54-dependent transcriptional activator of Pseudomonas putida.
    J Mol Biol. 1996 May 17;258(4):575-87 PMID: 8636993
  39. Signal sensing by sigma 54-dependent regulators: derepression as a control mechanism.
    Mol Microbiol. 1996 Feb;19(3):409-16 PMID: 8830233
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.1.41-45.1997
Published
1997-01-00
Pages
41-5
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178659
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.179.1.41-45.1997 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2