Comparison of the cleavage profiles of oligonucleotide duplexes with or without phosphorothioate linkages by using a chemical nuclease probe.
A manganese porphyrin complex, Mn-TMPyP, associated with KHSO(5) is a chemical nuclease able to selectively recognize the minor groove of three consecutive AT base pairs of DNA and to mediate very precise cleavage chemistry at that particular site. This specific recognition and cleavage were used to probe the accessibility of the minor groove of DNA duplexes composed of one phosphodiester strand and one phosphorothioate strand. The cleavage of 5'-GCAAAAGC/5'-GCTTTTGC duplexes by Mn-TMPyP/KHSO(5) was monitored by HPLC coupled to electrospray mass analysis. Each single strand was synthesized with all-phosphate, all- Rp-phosphorothioate and all- Sp-phosphorothioate internucleotide bonds. We found that the manganese porphyrin was able to recognize its favorite (AT)(3)-box binding site within the heteroduplexes, as in the case of natural DNA. Molecular modeling studies on the interactions of the reactive porphyrin manganese-oxo species with both types of duplexes confirmed the experimental data.
Chworos
A
A
Laboratoire de Chimie de Coordination du CNRS, 205 route de Narbonne, 31077 cedex 4, Toulouse, France.
Arnaud
P
P
Zakrzewska
K
K
Guga
P
P
Pratviel
G
G
Stec
W
W
Meunier
B
B
D001665:Binding Sites; D003851:Deoxyribonucleases / Q000737:chemistry*; D006868:Hydrolysis; D015335:Molecular Probes / Q000737:chemistry; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry*; D017553:Phosphorus Compounds / Q000737:chemistry*; D013873:Thionucleotides / Q000737:chemistry*
D015335:Molecular Probes; D009692:Nucleic Acid Heteroduplexes; D017553:Phosphorus Compounds; D013873:Thionucleotides; D003851:Deoxyribonucleases