Home LiteratureArticle Details
PMID: 7568230 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Structures of the apo- and the metal ion-activated forms of the diphtheria tox repressor from Corynebacterium diphtheriae.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 21 ·1995-10-10 ·Pages 9843-50

Schiering N, Tao X, Zeng H, Murphy JR, Petsko GA, Ringe D

Abstract

The diphtheria tox repressor (DtxR) of Corynebacterium diphtheriae plays a critical role in the regulation of diphtheria toxin expression and the control of other iron-sensitive genes. The crystal structures of apo-DtxR and of the metal ion-activated form of the repressor have been solved and used to identify motifs involved in DNA and metal ion binding. Residues involved in binding of the activated repressor to the diphtheria tox operator, glutamine 43, arginine 47, and arginine 50, were located and confirmed by site-directed mutagenesis. Previous biochemical and genetic data can be explained in terms of these structures. Conformational differences between apo- and Ni-DtxR are discussed with regard to the mechanism of action of this repressor.

MeSH Terms
Amino Acid Sequence Apoproteins/chemistry,genetics Bacterial Proteins/chemistry,genetics Binding Sites Corynebacterium diphtheriae/chemistry Crystallography, X-Ray DNA-Binding Proteins/chemistry,genetics Helix-Turn-Helix Motifs Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Nickel/chemistry Protein Binding Protein Conformation Repressor Proteins/chemistry,genetics Sequence Homology, Amino Acid
Chemicals
Apoproteins Bacterial Proteins DNA-Binding Proteins DtxR protein, Corynebacterium diphtheriae Repressor Proteins Nickel
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Schiering N
Department of Chemistry, Brandeis University, Waltham, MA 02154, USA.
Tao X
Zeng H
Murphy J R
Petsko G A
Ringe D
References (34)
34 references, click to expand
  1. Binding of the metalloregulatory protein DtxR to the diphtheria tox operator requires a divalent heavy metal ion and protects the palindromic sequence from DNase I digestion.
    J Biol Chem. 1992 Oct 25;267(30):21761-4 PMID: 1400485
  2. Three-dimensional structure of the diphtheria toxin repressor in complex with divalent cation co-repressors.
    Structure. 1995 Jan 15;3(1):87-100 PMID: 7743135
  3. The structure of the Antennapedia homeodomain determined by NMR spectroscopy in solution: comparison with prokaryotic repressors.
    Cell. 1989 Nov 3;59(3):573-80 PMID: 2572329
  4. SQUASH - combining constraints for macromolecular phase refinement and extension.
    Acta Crystallogr D Biol Crystallogr. 1993 Jan 1;49(Pt 1):213-22 PMID: 15299562
  5. Crystallization and preliminary X-ray studies of the diphtheria Tox repressor from Corynebacterium diphtheriae.
    J Mol Biol. 1994 Dec 16;244(5):654-6 PMID: 7990147
  6. 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
  7. Improved methods for building protein models in electron density maps and the location of errors in these models.
    Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9 PMID: 2025413
  8. Conformation of polypeptides and proteins.
    Adv Protein Chem. 1968;23:283-438 PMID: 4882249
  9. The helix-turn-helix DNA binding motif.
    J Biol Chem. 1989 Feb 5;264(4):1903-6 PMID: 2644244
  10. Resolution of phase ambiguity in macromolecular crystallography.
    Methods Enzymol. 1985;115:90-112 PMID: 4079800
  11. Escherichia coli biotin holoenzyme synthetase/bio repressor crystal structure delineates the biotin- and DNA-binding domains.
    Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9257-61 PMID: 1409631
  12. Repressor structure and the mechanism of positive control.
    Cell. 1983 Feb;32(2):319-25 PMID: 6218886
  13. DNA relationships among some tox-bearing corynebacteriophages.
    Infect Immun. 1985 Sep;49(3):679-84 PMID: 2993167
  14. A structural taxonomy of DNA-binding domains.
    Nature. 1991 Oct 24;353(6346):715-9 PMID: 1944532
  15. Cysteine-102 is positioned in the metal binding activation site of the Corynebacterium diphtheriae regulatory element DtxR.
    Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8524-8 PMID: 8378326
  16. Iron, DtxR, and the regulation of diphtheria toxin expression.
    Mol Microbiol. 1994 Oct;14(2):191-7 PMID: 7830565
  17. Determination of the minimal essential nucleotide sequence for diphtheria tox repressor binding by in vitro affinity selection.
    Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9646-50 PMID: 7937822
  18. Transition metals in control of gene expression.
    Science. 1993 Aug 6;261(5122):715-25 PMID: 8342038
  19. Transition metal ion activation of DNA binding by the diphtheria tox repressor requires the formation of stable homodimers.
    Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6803-7 PMID: 7624323
  20. Protein-DNA recognition.
    Annu Rev Biochem. 1984;53:293-321 PMID: 6236744
  21. Structure of the lambda complex at 2.5 A resolution: details of the repressor-operator interactions.
    Science. 1988 Nov 11;242(4880):893-9 PMID: 3187530
  22. Crystal structure of a CAP-DNA complex: the DNA is bent by 90 degrees.
    Science. 1991 Aug 30;253(5023):1001-7 PMID: 1653449
  23. Crystal structure of trp repressor/operator complex at atomic resolution.
    Nature. 1988 Sep 22;335(6188):321-9 PMID: 3419502
  24. Role of iron in regulation of virulence genes.
    Clin Microbiol Rev. 1993 Apr;6(2):137-49 PMID: 8472246
  25. Recognition of a DNA operator by the repressor of phage 434: a view at high resolution.
    Science. 1988 Nov 11;242(4880):899-907 PMID: 3187531
  26. Specific binding of the diphtheria tox regulatory element DtxR to the tox operator requires divalent heavy metal ions and a 9-base-pair interrupted palindromic sequence.
    Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5897-901 PMID: 1631071
  27. A method to identify protein sequences that fold into a known three-dimensional structure.
    Science. 1991 Jul 12;253(5016):164-70 PMID: 1853201
  28. Atomic structure of the actin:DNase I complex.
    Nature. 1990 Sep 6;347(6288):37-44 PMID: 2395459
  29. Iron-dependent regulation of diphtheria toxin and siderophore expression by the cloned Corynebacterium diphtheriae repressor gene dtxR in C. diphtheriae C7 strains.
    Infect Immun. 1991 Jun;59(6):1899-904 PMID: 1828057
  30. DNA sequences and characterization of dtxR alleles from Corynebacterium diphtheriae PW8(-), 1030(-), and C7hm723(-).
    J Bacteriol. 1992 Feb;174(4):1268-72 PMID: 1735717
  31. Crystal structure of LacI member, PurR, bound to DNA: minor groove binding by alpha helices.
    Science. 1994 Nov 4;266(5186):763-70 PMID: 7973627
  32. Engineered metal-binding proteins: purification to protein folding.
    Science. 1991 Jun 28;252(5014):1796-7 PMID: 1648261
  33. Characterization of mutations that inactivate the diphtheria toxin repressor gene (dtxR).
    Infect Immun. 1994 May;62(5):1600-8 PMID: 8168920
  34. Analysis of diphtheria toxin repressor-operator interactions and characterization of a mutant repressor with decreased binding activity for divalent metals.
    Mol Microbiol. 1993 Jul;9(1):173-81 PMID: 8412663
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.92.21.9843
Published
1995-10-10
Pages
9843-50
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC40899
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.92.21.9843 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: [email protected]

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