DNA footprinting with the hydroxyl radical.
The Fenton reaction of iron(II) EDTA with hydrogen peroxide, performed in the presence of ascorbate ion, has proven to be useful as a probe of structure in DNA systems. Two aspects of this chemistry are discussed: the identity of the active DNA cleaving agent produced by this reagent, and the application of the Fenton reaction to the determination of the structure of the Holliday junction, the four-stranded DNA molecule that is a key intermediate in recombination. The cleavage pattern of the Holliday junction has pseudo-twofold symmetry, putting important constraints on possible structures.
Tullius
T D
TD
Department of Chemistry, Johns Hopkins University, Baltimore, Maryland 21218.
D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004249:DNA Damage; D004279:DNA, Viral / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005609:Free Radicals; D005720:Gamma Rays; D006878:Hydroxides; D017665:Hydroxyl Radical; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012097:Repressor Proteins / Q000737:chemistry
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D005609:Free Radicals; D006878:Hydroxides; D012097:Repressor Proteins; D017665:Hydroxyl Radical