Monitoring ssDNA binding to the DnaB helicase from $Helicobacter\ pylori$ by solid-state NMR spectroscopy - Groupe Pathogénie bactérienne et réponses cellulaires / Integrated Imaging of Stress Response Group (IBS-I2SR) Accéder directement au contenu
Article Dans Une Revue Angewandte Chemie International Edition Année : 2016

Monitoring ssDNA binding to the DnaB helicase from $Helicobacter\ pylori$ by solid-state NMR spectroscopy

Résumé

DnaB helicases are bacterial, ATP-driven enzymes that unwind double-stranded DNA during DNA replication. Herein, we study the sequential binding of the "non-hydrolysable" ATP analogue AMP-PNP and of single-stranded (ss) DNA to the dodecameric DnaB helicase from Helicobacter pylori using solid-state NMR. Phosphorus cross-polarization experiments monitor the binding of AMP-PNP and DNA to the helicase. 13 C chemical-shift perturbations (CSPs) are used to detect conformational changes in the protein upon binding. The helicase switches upon AMP-PNP addition into a conformation apt for ssDNA binding, and AMP-PNP is hydrolyzed and released upon binding of ssDNA. Our study sheds light on the conformational changes which are triggered by the interaction with AMP-PNP and are needed for ssDNA binding of H. pylori DnaB in vitro. They also demonstrate the level of detail solid-state NMR can provide for the characterization of protein-DNA interactions and the interplay with ATP or its analogues.
Fichier principal
Vignette du fichier
Wiegand_AngChem_2016.pdf (3.57 Mo) Télécharger le fichier
Origine Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-01430549 , version 1 (07-06-2024)

Identifiants

Citer

Thomas Wiegand, Riccardo Cadalbert, Carole Gardiennet, Joanna Timmins, Laurent Terradot, et al.. Monitoring ssDNA binding to the DnaB helicase from $Helicobacter\ pylori$ by solid-state NMR spectroscopy. Angewandte Chemie International Edition, 2016, 55 (45), pp.14164 - 14168. ⟨10.1002/anie.201607295⟩. ⟨hal-01430549⟩
128 Consultations
2 Téléchargements

Altmetric

Partager

Gmail Mastodon Facebook X LinkedIn More