Cited by 57Diphtheria, Corynebacterium, and TetanusBacterial Genetics and BiotechnologyProtein Structure and Dynamics

Mechanism of metal ion activation of the diphtheria toxin repressor DtxR

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D'Aquino, J. Alejandro, Tetenbaum-Novatt, Jaclyn, White, Andre, Berkovitch, Fred, Ringe, Dagmar·2005·Proceedings of the National Academy of Sciences

Abstract

The diphtheria toxin repressor (DtxR) is a metal ion-activated transcriptional regulator that has been linked to the virulence of Corynebacterium diphtheriae . Structure determination has shown that there are two metal ion binding sites per repressor monomer, and site-directed mutagenesis has demonstrated that binding site 2 (primary) is essential for recognition of the target DNA repressor, leaving the role of binding site 1 (ancillary) unclear. Calorimetric techniques have demonstrated that although binding site 1 (ancillary) has high affinity for metal ion with a binding constant of 2 × 10 –7 , binding site 2 (primary) is a low-affinity binding site with a binding constant of 6.3 × 10 –4 . These two binding sites act in an independent fashion, and their contribution can be easily dissected by traditional mutational analysis. Our results clearly demonstrate that binding site 1 (ancillary) is the first one to be occupied during metal ion activation, playing a critical role in stabilization of the repressor. In addition, structural data obtained for the mutants Ni-DtxR(H79A,C102D), reported here, and the previously reported DtxR(H79A) have allowed us to propose a mechanism of metal activation for DtxR.

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Crossref · 10.1073/pnas.0500908102

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Authors
D'Aquino, J. A., Tetenbaum-Novatt, J., White, A., Berkovitch, F., & Ringe, D.
Affiliations
  • Departments of Chemistry and Biochemistry and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02454; and Department of Medicinal Chemistry, Boehringer Ingelheim Pharmaceuticals, Ridgefield, CT 06877
  • Boehringer Ingelheim (United States) (US)
Corresponding author
Ringe, Dagmar
Venue
Proceedings of the National Academy of Sciences
Published
2005-12-13
Volume
102
Issue
51
Pages
18408-18413
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
OpenAlex
W1982757640
Cited by
57
References
38
Citations by year
2026: 3 · 2025: 1 · 2024: 5 · 2023: 2 · 2022: 2 · 2021: 1 · 2020: 2 · 2019: 2 · 2017: 3 · 2016: 2 · 2015: 3 · 2014: 1 · 2013: 6 · 2012: 6
FWCI
1.62
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Science Foundation (GM68762, 0070319)
  • School of Medicine, Boston University
  • National Institutes of Health (0070319, 3p50gm068762-07s2, 1s10rr010493-01, GM68762)
  • National Institute of General Medical Sciences (GM 68762, P50-GM068762)
  • National Institute of Allergy and Infectious Diseases (AI-21628, R01AI021628)
  • National Center for Research Resources (P41-RR-10888, S10 RR-10493, P41RR010888)
  • Brookhaven National Laboratory
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Diphtheria, Corynebacterium, and Tetanus
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
DtxR, metal ion activation, metal binding sites, transcriptional regulation, Corynebacterium diphtheriae, virulence, protein stability, DNA binding, nickel binding
MeSH terms
Bacterial Proteins, Binding Sites, Calorimetry, Corynebacterium diphtheriae, Cysteine, DNA-Binding Proteins, Histidine, Ligands, Nickel, Protein Denaturation, Temperature, Protein Folding, Crystallography, X-Ray

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