Tetracycline repressor allostery does not depend on divalent metal recognition

Sebastiaan Werten, Daniela Dalm, Gottfried Julius Palm, Christopher Cornelius Grimm, Winfried Hinrichs

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Genes that render bacteria resistant to tetracycline-derived antibiotics are tightly regulated by repressors of the TetR family. In their physiologically relevant, magnesium-complexed form, tetracyclines induce allosteric rearrangements in the TetR homodimer, leading to its release from the promoter and derepression of transcription. According to earlier crystallographic work, recognition of the tetracycline-associated magnesium ion by TetR is crucial and triggers the allosteric cascade. Nevertheless, the derivative 5a,6-anhydrotetracycline, which shows an increased affinity for TetR, causes promoter release even in the absence of magnesium. To resolve this paradox, it has been proposed that metal-free 5a,6-anhydrotetracycline acts via an exceptional, conformationally different induction mode that circumvents the normal magnesium requirement. We have tested this hypothesis by determining crystal structures of TetR-5a,6-anhydrotetracycline complexes in the presence of magnesium, ethylenediaminetetraacetic acid, or high concentrations of potassium. Analysis of these three structures reveals that, irrespective of the metal, the effects of 5a,6-anhydrotetracycline binding are indistinguishable from those of canonical induction by other tetracyclines. Together with a close scrutiny of the earlier evidence of a metal-triggered mechanism, these results demonstrate that magnesium recognition per se is not a prerequisite for tetracycline repressor allostery.

Original languageEnglish (US)
Pages (from-to)7990-7998
Number of pages9
JournalBiochemistry
Volume53
Issue number50
DOIs
StatePublished - Dec 23 2014
Externally publishedYes

Fingerprint

Tetracycline
Magnesium
Metals
Tetracyclines
Transcription
Edetic Acid
Bacteria
Potassium
Genes
Crystal structure
Ions
Anti-Bacterial Agents
Derivatives
4-epianhydrotetracycline

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)

Cite this

Werten, S., Dalm, D., Palm, G. J., Grimm, C. C., & Hinrichs, W. (2014). Tetracycline repressor allostery does not depend on divalent metal recognition. Biochemistry, 53(50), 7990-7998. https://doi.org/10.1021/bi5012805

Tetracycline repressor allostery does not depend on divalent metal recognition. / Werten, Sebastiaan; Dalm, Daniela; Palm, Gottfried Julius; Grimm, Christopher Cornelius; Hinrichs, Winfried.

In: Biochemistry, Vol. 53, No. 50, 23.12.2014, p. 7990-7998.

Research output: Contribution to journalArticle

Werten, S, Dalm, D, Palm, GJ, Grimm, CC & Hinrichs, W 2014, 'Tetracycline repressor allostery does not depend on divalent metal recognition', Biochemistry, vol. 53, no. 50, pp. 7990-7998. https://doi.org/10.1021/bi5012805
Werten S, Dalm D, Palm GJ, Grimm CC, Hinrichs W. Tetracycline repressor allostery does not depend on divalent metal recognition. Biochemistry. 2014 Dec 23;53(50):7990-7998. https://doi.org/10.1021/bi5012805
Werten, Sebastiaan ; Dalm, Daniela ; Palm, Gottfried Julius ; Grimm, Christopher Cornelius ; Hinrichs, Winfried. / Tetracycline repressor allostery does not depend on divalent metal recognition. In: Biochemistry. 2014 ; Vol. 53, No. 50. pp. 7990-7998.
@article{f42cbe2e6c624111914bde89b919a90d,
title = "Tetracycline repressor allostery does not depend on divalent metal recognition",
abstract = "Genes that render bacteria resistant to tetracycline-derived antibiotics are tightly regulated by repressors of the TetR family. In their physiologically relevant, magnesium-complexed form, tetracyclines induce allosteric rearrangements in the TetR homodimer, leading to its release from the promoter and derepression of transcription. According to earlier crystallographic work, recognition of the tetracycline-associated magnesium ion by TetR is crucial and triggers the allosteric cascade. Nevertheless, the derivative 5a,6-anhydrotetracycline, which shows an increased affinity for TetR, causes promoter release even in the absence of magnesium. To resolve this paradox, it has been proposed that metal-free 5a,6-anhydrotetracycline acts via an exceptional, conformationally different induction mode that circumvents the normal magnesium requirement. We have tested this hypothesis by determining crystal structures of TetR-5a,6-anhydrotetracycline complexes in the presence of magnesium, ethylenediaminetetraacetic acid, or high concentrations of potassium. Analysis of these three structures reveals that, irrespective of the metal, the effects of 5a,6-anhydrotetracycline binding are indistinguishable from those of canonical induction by other tetracyclines. Together with a close scrutiny of the earlier evidence of a metal-triggered mechanism, these results demonstrate that magnesium recognition per se is not a prerequisite for tetracycline repressor allostery.",
author = "Sebastiaan Werten and Daniela Dalm and Palm, {Gottfried Julius} and Grimm, {Christopher Cornelius} and Winfried Hinrichs",
year = "2014",
month = "12",
day = "23",
doi = "10.1021/bi5012805",
language = "English (US)",
volume = "53",
pages = "7990--7998",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "50",

}

TY - JOUR

T1 - Tetracycline repressor allostery does not depend on divalent metal recognition

AU - Werten, Sebastiaan

AU - Dalm, Daniela

AU - Palm, Gottfried Julius

AU - Grimm, Christopher Cornelius

AU - Hinrichs, Winfried

PY - 2014/12/23

Y1 - 2014/12/23

N2 - Genes that render bacteria resistant to tetracycline-derived antibiotics are tightly regulated by repressors of the TetR family. In their physiologically relevant, magnesium-complexed form, tetracyclines induce allosteric rearrangements in the TetR homodimer, leading to its release from the promoter and derepression of transcription. According to earlier crystallographic work, recognition of the tetracycline-associated magnesium ion by TetR is crucial and triggers the allosteric cascade. Nevertheless, the derivative 5a,6-anhydrotetracycline, which shows an increased affinity for TetR, causes promoter release even in the absence of magnesium. To resolve this paradox, it has been proposed that metal-free 5a,6-anhydrotetracycline acts via an exceptional, conformationally different induction mode that circumvents the normal magnesium requirement. We have tested this hypothesis by determining crystal structures of TetR-5a,6-anhydrotetracycline complexes in the presence of magnesium, ethylenediaminetetraacetic acid, or high concentrations of potassium. Analysis of these three structures reveals that, irrespective of the metal, the effects of 5a,6-anhydrotetracycline binding are indistinguishable from those of canonical induction by other tetracyclines. Together with a close scrutiny of the earlier evidence of a metal-triggered mechanism, these results demonstrate that magnesium recognition per se is not a prerequisite for tetracycline repressor allostery.

AB - Genes that render bacteria resistant to tetracycline-derived antibiotics are tightly regulated by repressors of the TetR family. In their physiologically relevant, magnesium-complexed form, tetracyclines induce allosteric rearrangements in the TetR homodimer, leading to its release from the promoter and derepression of transcription. According to earlier crystallographic work, recognition of the tetracycline-associated magnesium ion by TetR is crucial and triggers the allosteric cascade. Nevertheless, the derivative 5a,6-anhydrotetracycline, which shows an increased affinity for TetR, causes promoter release even in the absence of magnesium. To resolve this paradox, it has been proposed that metal-free 5a,6-anhydrotetracycline acts via an exceptional, conformationally different induction mode that circumvents the normal magnesium requirement. We have tested this hypothesis by determining crystal structures of TetR-5a,6-anhydrotetracycline complexes in the presence of magnesium, ethylenediaminetetraacetic acid, or high concentrations of potassium. Analysis of these three structures reveals that, irrespective of the metal, the effects of 5a,6-anhydrotetracycline binding are indistinguishable from those of canonical induction by other tetracyclines. Together with a close scrutiny of the earlier evidence of a metal-triggered mechanism, these results demonstrate that magnesium recognition per se is not a prerequisite for tetracycline repressor allostery.

UR - http://www.scopus.com/inward/record.url?scp=84919740397&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84919740397&partnerID=8YFLogxK

U2 - 10.1021/bi5012805

DO - 10.1021/bi5012805

M3 - Article

VL - 53

SP - 7990

EP - 7998

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 50

ER -