title: Solid-state NMR chemical-shift perturbations indicate domain reorientation of the DnaG primase in the primosome of Helicobacter pylori.
abstract: We here investigate the interactions between the DnaB helicase and the C-terminal domain of the corresponding DnaG primase of Helicobacter pylori using solid-state NMR. The difficult crystallization of this 387 kDa complex, where the two proteins interact in a six to three ratio, is circumvented by simple co-sedimentation of the two proteins directly into the MAS-NMR rotor. While the amount of information that can be extracted from such a large protein is still limited, we can assign a number of amino-acid residues experiencing significant chemical-shift perturbations upon helicase-primase complex formation. The location of these residues is used as a guide to model the interaction interface between the two proteins in the complex. Chemical-shift perturbations also reveal changes at the interaction interfaces of the hexameric HpDnaB assembly on HpDnaG binding. A structural model of the complex that explains the experimental findings is obtained.
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France.|CNRS, CRM2, UMR 7036, Université de Lorraine, 54506, Vandoeuvre-lès-Nancy, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Gardiennet
authors.affiliation: Physical Chemistry, ETH Zurich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wiegand
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Bazin
authors.affiliation: Physical Chemistry, ETH Zurich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Cadalbert
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France.
authors.forename: Britta
authors.initials: B
authors.lastname: Kunert
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Lacabanne
authors.affiliation: Institut de Biologie Structurale (IBS), CNRS, IBS; CEA, IBS, Université Grenoble Alpes, 38044, Grenoble, France.
authors.forename: Irina
authors.initials: I
authors.lastname: Gutsche
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France. l.terradot@ibcp.fr.
authors.forename: Laurent
authors.initials: L
authors.lastname: Terradot
authors.affiliation: Physical Chemistry, ETH Zurich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland. beme@ethz.ch.
authors.forename: Beat H
authors.initials: BH
authors.lastname: Meier
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France. a.bockmann@ibcp.fr.
authors.forename: Anja
authors.initials: A
authors.lastname: Böckmann
mesh_terms: D019915:DNA Primase / Q000737:chemistry*; D016480:Helicobacter pylori / Q000201:enzymology*; D019906:Nuclear Magnetic Resonance, Biomolecular; D000072417:Protein Domains
chemical_list: D019915:DNA Primase
keywords: DnaB; DnaG; Primosome; Protein complexes; Sediments; Solid-state NMR