Radiolytic footprinting. .beta. Rays, .gamma. photons, and fast neutrons probe DNA-protein interactions

Ionizing radiations induce numerous damages in DNA, especially strand breaks. The hydroxyl radical OH., produced by the radiolysis of water, is mainly responsible for this effect. The fact that strand breakage occurs at all nucleotides and that bound proteins may locally radioprotect DNA at the bind...

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Bibliographic Details
Published in:Biochemistry (Easton) 1993-03, Vol.32 (8), p.2104-2110
Main Authors: Franchet-Beuzit, Jenny, Spotheim-Maurizot, Melanie, Sabattier, Roland, Blazy-Baudras, Bernadette, Charlier, Michel
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Beta Particles
Biological and medical sciences
Carrier Proteins - metabolism
Carrier Proteins - radiation effects
Chickens
Cyclic AMP Receptor Protein
DNA - metabolism
DNA - radiation effects
DNA Damage
DNA, Bacterial - genetics
DNA, Bacterial - metabolism
DNA, Bacterial - radiation effects
Escherichia coli - genetics
Escherichia coli - metabolism
Free Radicals
Fundamental and applied biological sciences. Psychology
Gamma Rays
Hydroxides
Hydroxyl Radical
Interactions. Associations
Intermolecular phenomena
Lac Operon - radiation effects
Molecular biophysics
Molecular Sequence Data
Neutrons
Nucleic Acid Conformation
Nucleosomes - metabolism
Plasmids - radiation effects
Receptors, Cyclic AMP - metabolism
Receptors, Cyclic AMP - radiation effects
Repressor Proteins - metabolism
Repressor Proteins - radiation effects
Restriction Mapping
Ultraviolet Rays
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Summary:Ionizing radiations induce numerous damages in DNA, especially strand breaks. The hydroxyl radical OH., produced by the radiolysis of water, is mainly responsible for this effect. The fact that strand breakage occurs at all nucleotides and that bound proteins may locally radioprotect DNA at the binding site lead us to develop a radiolytic footprinting method to study DNA-protein interactions. Three different radiations were used: beta rays, gamma photons, and fast neutrons. In order to validate this technique, three well-known interaction systems were tested: the lac repressor-lac operator of Escherichia coli, the cyclic AMP receptor protein (CRP) of E. coli and its specific site in the lac regulation region, and the core nucleosome. Radiolytic footprinting gives results similar to those obtained by more classical probes: DNase I, complexes of orthophenanthroline (OP) and copper, complexes of ethylenediaminetetraacetate ion (EDTA) and iron, and UV light. For the same system (lac repressor), irradiation with either gamma photons or fast neutrons gives identical results.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00059a031