title: The RecB nuclease domain binds to RecA-DNA filaments: implications for filament loading.
abstract: The E. coli RecBCD enzyme facilitates the loading of RecA onto single-stranded DNA produced by the combined helicase/nuclease activity of RecBCD. The nuclease domain of RecB protein, RecB(nuc), has been previously shown to bind RecA. Surprisingly, RecB(nuc) also binds to phage and eukaryotic homologs of RecA, leading to the suggestion that RecB(nuc) interacts with the polymerization motif that is present in all three proteins. This mode of interaction could only be with monomeric RecA, as this motif would be buried in filaments. We show that RecB(nuc) binds extensively to the outside of RecA-DNA filaments. Three-dimensional reconstructions suggest that RecB(nuc) binds to the ATP-binding core of RecA, with a displacement of the C-terminal domain of RecA. Solution experiments confirm that the interaction of RecB(nuc) is only with the RecA core. Since the RecA C-terminal domain has been shown to be regulatory, the interaction observed may be part of the loading mechanism where RecB displaces the RecA C-terminal domain and activates a RecA monomer for polymerization.
authors.affiliation: Cancer Research UK Clare Hall Laboratories, The London Research Institute, Herts, U.K.
authors.forename: Debora
authors.initials: D
authors.lastname: Lucarelli
authors.forename: Ying A
authors.initials: YA
authors.lastname: Wang
authors.forename: Vitold E
authors.initials: VE
authors.lastname: Galkin
authors.forename: Xiong
authors.initials: X
authors.lastname: Yu
authors.forename: Dale B
authors.initials: DB
authors.lastname: Wigley
authors.forename: Edward H
authors.initials: EH
authors.lastname: Egelman
mesh_terms: D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D043211:Exodeoxyribonuclease V / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011938:Rec A Recombinases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D004247:DNA; D011938:Rec A Recombinases; D043211:Exodeoxyribonuclease V; C468322:exodeoxyribonuclease V, E coli