Genetic switching by the Lac repressor is based on two-state Monod–Wyman–Changeux allostery

Julija Romanuka; Gert e. Folkers; Manuel Gnida; Lidija Kovačič; Hans Wienk; Robert Kaptein; Rolf Boelens

doi:10.1073/pnas.2311240120

Genetic switching by the Lac repressor is based on two-state Monod–Wyman–Changeux allostery

Julija Romanuka
, Gert e. Folkers
, Manuel Gnida
, Lidija Kovačič
, Hans Wienk
, Robert Kaptein
, Rolf Boelens^*

^*Corresponding author for this work

NMR Spectroscopy, Bijvoet Centre for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The Netherlands

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

High-resolution NMR spectroscopy enabled us to characterize allosteric transitions between various functional states of the dimeric Escherichia coli Lac repressor. In the absence of ligands, the dimer exists in a dynamic equilibrium between DNA-bound and inducer-bound conformations. Binding of either effector shifts this equilibrium toward either bound state. Analysis of the ternary complex between repressor, operator DNA, and inducer shows how adding the inducer results in allosteric changes that disrupt the interdomain contacts between the inducer binding and DNA binding domains and how this in turn leads to destabilization of the hinge helices and release of the Lac repressor from the operator. Based on our data, the allosteric mechanism of the induction process is in full agreement with the well-known Monod–Wyman–Changeux model.

Original language	English
Article number	2311240120
Pages (from-to)	1-8
Number of pages	8
Journal	Proceedings of the National Academy of Sciences
Volume	120
Issue number	49
DOIs	https://doi.org/10.1073/pnas.2311240120
Publication status	Published - 5 Dec 2023

Bibliographical note

Publisher Copyright:
Copyright © 2023 the Author(s). Published by PNAS. This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

Funding

ACKNOWLEDGMENTS. Dr. J. Grinstead is acknowledged for critical reading of the manuscript.S.J.de Vries is acknowledged for providing scripts for theanalysis of some chemical shift data. R.K. and R.B. were supported by the Netherlands Foundation for Chemical Research (NWO-CW, 175.107.301.10; 700.52.303; and 700.53.103). R.B. was supported by the European Commission (project 031220, Spine2-complexes; project RII3-026145,EU-NMR; project 261863,BioNMR; and project 228461, East-NMR). M.G. was supported by a stipend of the Deutsche Forschungsgemeinschaft (DFG, project 5444273).

Funders	Funder number
Netherlands Foundation for Chemical Research
European Commission	RII3-026145, 228461, 261863, 031220
Deutsche Forschungsgemeinschaft	5444273
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	175.107.301.10, 700.53.103, 700.52.303

Keywords

NMR
allostery
gene regulation
repressor
structural biology

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10.1073/pnas.2311240120

romanuka-et-al-2023-genetic-switching-by-the-lac-repressor-is-based-on-two-state-monod-wyman-changeux-allosteryFinal published version, 1.94 MBLicence: Taverne

Cite this

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title = "Genetic switching by the Lac repressor is based on two-state Monod–Wyman–Changeux allostery",

abstract = "High-resolution NMR spectroscopy enabled us to characterize allosteric transitions between various functional states of the dimeric Escherichia coli Lac repressor. In the absence of ligands, the dimer exists in a dynamic equilibrium between DNA-bound and inducer-bound conformations. Binding of either effector shifts this equilibrium toward either bound state. Analysis of the ternary complex between repressor, operator DNA, and inducer shows how adding the inducer results in allosteric changes that disrupt the interdomain contacts between the inducer binding and DNA binding domains and how this in turn leads to destabilization of the hinge helices and release of the Lac repressor from the operator. Based on our data, the allosteric mechanism of the induction process is in full agreement with the well-known Monod–Wyman–Changeux model.",

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author = "Julija Romanuka and Folkers, \{Gert e.\} and Manuel Gnida and Lidija Kova{\v c}i{\v c} and Hans Wienk and Robert Kaptein and Rolf Boelens",

note = "Publisher Copyright: Copyright {\textcopyright} 2023 the Author(s). Published by PNAS. This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).",

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AU - Romanuka, Julija

AU - Folkers, Gert e.

AU - Gnida, Manuel

AU - Kovačič, Lidija

AU - Wienk, Hans

AU - Kaptein, Robert

AU - Boelens, Rolf

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AB - High-resolution NMR spectroscopy enabled us to characterize allosteric transitions between various functional states of the dimeric Escherichia coli Lac repressor. In the absence of ligands, the dimer exists in a dynamic equilibrium between DNA-bound and inducer-bound conformations. Binding of either effector shifts this equilibrium toward either bound state. Analysis of the ternary complex between repressor, operator DNA, and inducer shows how adding the inducer results in allosteric changes that disrupt the interdomain contacts between the inducer binding and DNA binding domains and how this in turn leads to destabilization of the hinge helices and release of the Lac repressor from the operator. Based on our data, the allosteric mechanism of the induction process is in full agreement with the well-known Monod–Wyman–Changeux model.

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KW - gene regulation

KW - repressor

KW - structural biology

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DO - 10.1073/pnas.2311240120

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Genetic switching by the Lac repressor is based on two-state Monod–Wyman–Changeux allostery

Abstract

Bibliographical note

Funding

Keywords

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