title: Expression, purification, crystallization and preliminary X-ray diffraction analysis of the novel modular DNA-binding protein BurrH in its apo form and in complex with its target DNA.
abstract: Different genome-editing strategies have fuelled the development of new DNA-targeting molecular tools allowing precise gene modifications. Here, the expression, purification, crystallization and preliminary X-ray diffraction of BurrH, a novel DNA-binding protein from Burkholderia rhizoxinica, are reported. Crystallization experiments of BurrH in its apo form and in complex with its target DNA yielded crystals suitable for X-ray diffraction analysis. The crystals of the apo form belonged to the primitive hexagonal space group P3(1) or its enantiomorph P3(2), with unit-cell parameters a = b = 73.28, c = 268.02 Å, α = β = 90, γ = 120°. The BurrH-DNA complex crystallized in the monoclinic space group P2(1), with unit-cell parameters a = 70.15, b = 95.83, c = 76.41 Å, α = γ = 90, β = 109.51°. The self-rotation function and the Matthews coefficient suggested the presence of two protein molecules per asymmetric unit in the apo crystals and one protein-DNA complex in the monoclinic crystals. The crystals diffracted to resolution limits of 2.21 and 2.65 Å, respectively, using synchrotron radiation.
authors.affiliation: Macromolecular Crystallography Group, Structural Biology and Biocomputing Programme, Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid, Spain.
authors.forename: Stefano
authors.initials: S
authors.lastname: Stella
authors.affiliation: Macromolecular Crystallography Group, Structural Biology and Biocomputing Programme, Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Molina
authors.affiliation: Cellectis, 8 Rue de la Croix Jarry, 75013 Paris, France.
authors.forename: Claudia
authors.initials: C
authors.lastname: Bertonatti
authors.affiliation: Cellectis, 8 Rue de la Croix Jarry, 75013 Paris, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Juillerrat
authors.affiliation: Macromolecular Crystallography Group, Structural Biology and Biocomputing Programme, Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid, Spain.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Montoya
mesh_terms: D001059:Apoproteins / Q000737:chemistry* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D019117:Burkholderia / Q000201:enzymology*; D003460:Crystallization; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D014961:X-Ray Diffraction
chemical_list: D001059:Apoproteins; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D004247:DNA
keywords: Burkholderia rhizoxinica; BurrH; gene targeting; protein–DNA interaction