title: The location of DNA in complexes of recA protein with double-stranded DNA. A neutron scattering study.
abstract: Purified recA protein is found as rodlike homopolymers, and it forms filamentous complexes with double-stranded DNA that are stable in the presence of ATP gamma S, a nonhydrolyzable analogue of ATP. The structure of these filaments has been described in some detail by electron microscopy. Here we confirm the mass per length of 6.5 recA/100 A in solution by small-angle neutron scattering and extend the analysis to homopolymers of recA protein, finding a mass per length of about 7 recA/100 A and a radial mass distribution (cross-sectional radius of gyration) significantly different for the two filaments. The models proposed so far for the structure of the complex have placed the DNA in the center of the filament. Here we verify this assumption using small-angle neutron scattering to locate the DNA in the complexes, exploiting the contrast variation method in D2O/H2O mixtures. Model calculations show that the natural contrast difference between DNA and protein is not sufficient to locate the DNA (which accounts for only 4.7% of the mass in the complex). When deuterated DNA is used, the contrast difference is enhanced, and model calculations and experiment then converge, indicating that the DNA is indeed near the axis of the complex.
authors.affiliation: Institut Laue-Langevin, Grenoble, France.
authors.forename: E
authors.initials: E
authors.lastname: DiCapua
authors.forename: M
authors.initials: M
authors.lastname: Schnarr
authors.forename: P A
authors.initials: PA
authors.lastname: Timmins
mesh_terms: D001665:Binding Sites; D004269:DNA, Bacterial; D003903:Deuterium; D004926:Escherichia coli; D008956:Models, Chemical; D015394:Molecular Structure; D009502:Neutrons; D011938:Rec A Recombinases; D012542:Scattering, Radiation
chemical_list: D004269:DNA, Bacterial; D003903:Deuterium; D011938:Rec A Recombinases