A new family of sequence-specific DNA-cleaving agents directed by triple-helical structures: benzopyridoindole-EDTA conjugates.
Sequence-specific DNA recognition can be achieved by oligonucleotides that bind to the major groove of oligopyrimidine x oligopurine sequences. These intermolecular structures could be used to modulate gene expression and to create new tools for molecular biology. Here we report the synthesis and biochemical characterization of triple helix-specific DNA cleaving reagents. It is based on the previously reported triplex-specific ligands, benzo[e]pyridoindole (BePI) and benzo[g]pyridoindole (BgPI), covalently attached to ethylenediaminotetraacetic acid (EDTA). In the presence of iron, a reducing agent and molecular oxygen, BgPI-EDTA x FeII but not BePI-EDTA x FeII induced a double-stranded cut in a plasmid DNA at the single site where a triplex-forming oligonucleotide binds. At single nucleotide resolution, it was found that upon triplex formation BePI-EDTA x FeII led to cleavage of the pyrimidine strand and protection of the purine strand. BgPI-EDTA x FeII cleaved both strands with similar efficiency. The difference in cleavage efficiency between the two conjugates was rationalized by the location of the EDTA x FeII moiety with respect to the grooves of DNA (major groove: BePI-EDTA x FeII, minor groove: BgPI-EDTA x FeII). This work paves the way to the development of a new class of triple helix directed DNA cleaving reagents. Such molecules will be of interest for sequence-specific DNA cleavage and for investigating triple-helical structures, such as H-DNA, which could play an important role in the control of gene expression in vivo.
Marchand
C
C
Laboratoire de Biophysique, UMR8646 CNRS-Muséum National d'Histoire Naturelle, INSERM U201, Paris, France.
Nguyen
C H
CH
Ward
B
B
Sun
J S
JS
Bisagni
E
E
Garestier
T
T
Hélène
C
C
D004247:DNA / Q000737:chemistry*; D004492:Edetic Acid / Q000737:chemistry*; D005786:Gene Expression Regulation / Q000187:drug effects; D007211:Indoles / Q000737:chemistry*; D007501:Iron / Q000737:chemistry; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry
D007211:Indoles; D009838:Oligodeoxyribonucleotides; D004247:DNA; D004492:Edetic Acid; D007501:Iron