DNA cleavage by hydroxy-salicylidene-ethylendiamine-iron complexes.
Bis(hydroxy)salen.Fe complexes were designed as self-activated chemical nucleases. The presence of a hy-droxyl group on the two salicylidene moieties serve to form a hydroquinone system cooperating with the iron redox system to facilitate spontaneous formation of free radicals. We compared the DNA binding and cleaving properties of the ortho -, meta- and para -(bishydroxy) salen.Fe complexes with that of the corresponding chelate lacking the hydroxyl groups. DNA melting temperature studies indicated that the para complex exhibits the highest affinity for DNA. In addition, this para compound was considerably more potent at cleaving supercoiled plasmid DNA than the regio-isomeric ortho - and meta -hydroxy-salen.Fe complexes, even in the absence of a reducing agent, such as dithiothreitol used to activate the metal complex. The DNA cleaving activity of the para isomer is both time and concentration dependent and the complexed iron atom is absolutely essential for the sequence uniform cleavage of DNA. From a mechanistic point of view, electron spin resonance measurements suggest that DNA contributes positively to the activation of the semi-quinone system and the production of ligand radical species responsible for subsequent strand scission in the absence of a reducing agent. The para -hydroxy-salen.Fe complex has been used for detecting sequence-specific drug-DNA interactions. Specific binding of Hoechst 33258 to AT sequences and chromomycin to GC sequences were shown. The para -bis(hydroxy)salen.Fe derivative complements the tool box of footprinting reagents which can be utilised to produce efficient cleavage of DNA.
Routier
S
S
Laboratoire de Chimie Organique Physique, URA CNRS 351, USTL Bât. C3, 59655 Villeneuve d'Ascq, France.
Vezin
H
H
Lamour
E
E
Bernier
J L
JL
Catteau
J P
JP
Bailly
C
C
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D006690:Bisbenzimidazole / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D002614:Chelating Agents / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018983:DNA Footprinting; D004278:DNA, Superhelical / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004229:Dithiothreitol / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005029:Ethylenediamines / Q000737:chemistry / Q000378:metabolism*; D005609:Free Radicals / Q000378:metabolism; D006900:Hydroxylation; D007364:Intercalating Agents / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D007536:Isomerism; D009691:Nucleic Acid Denaturation; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism; D019163:Reducing Agents / Q000378:metabolism; D012545:Schiff Bases / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
D002614:Chelating Agents; D004278:DNA, Superhelical; D005029:Ethylenediamines; D005609:Free Radicals; D007364:Intercalating Agents; D017382:Reactive Oxygen Species; D019163:Reducing Agents; D012545:Schiff Bases; D004247:DNA; C011452:disalicylaldehyde ethylenediamine; D007501:Iron; D004706:Endodeoxyribonucleases; D006690:Bisbenzimidazole; D004229:Dithiothreitol