title: Detection of DNA damage: effect of thymidine glycol residues on the thermodynamic, substrate and interfacial acoustic properties of oligonucleotide duplexes.
abstract: Thymidine glycol residues in DNA are biologically active oxidative molecular damage sites caused by ionizing radiation and other factors. One or two thymidine glycol residues were incorporated in 19- to 31-mer DNA fragments during automatic oligonucleotide synthesis. These oligonucleotide models were used to estimate the effect of oxidized thymidines on the thermodynamic, substrate and interfacial acoustic properties of DNA. UV-monitoring melting data revealed that modified residues in place of thymidines destabilize the DNA double helix by 8-22 degrees C, depending on the number of lesions, the length of oligonucleotide duplexes and their GC-content. The diminished hybridizing capacity of modified oligonucleotides is presumably due to the loss of aromaticity and elevated hydrophilicity of thymine glycol in comparison to the thymine base. According to circular dichroism (CD) data, the modified DNA duplexes retain B-form geometry, and the thymidine glycol residue introduces only local perturbations limited to the lesion site. The rate of DNA hydrolysis by restriction endonucleases R.MvaI, R.Bst2UI, R.MspR9I and R.Bme1390I is significantly decreased as the thymidine glycol is located in the central position of the double-stranded recognition sequences 5'-CC / WGG-3' (W = A, T) or 5'-CC / NGG-3' (N = A, T, G, C) adjacent to the cleavage site. On the other hand, the catalytic properties of enzymes R.Psp6I and R.BstSCI recognizing the similar sequence are not changed dramatically, since their cleavage site is separated from the point of modification by several base-pairs. Data obtained by gel-electrophoretic analysis of radioactive DNA substrates were confirmed by direct spectrophotometric assay developed by the authors. The effect of thymidine glycol was also observed on DNA hybridization at the surface of a thickness-shear mode acoustic wave device. A 1.9-fold decrease in the rate of duplex formation was noted for oligonucleotides carrying one or two thymidine glycol residues in relation to the unmodified analog.
authors.affiliation: Chemistry Department and A.N. Belozersky Institute of Physico-Chemical Biology, M. V. Lomonosov Moscow State University, Leninskije Gory, Moscow, 119 991, Russia.
authors.forename: F
authors.initials: F
authors.lastname: Yang
authors.forename: E
authors.initials: E
authors.lastname: Romanova
authors.forename: E
authors.initials: E
authors.lastname: Kubareva
authors.forename: N
authors.initials: N
authors.lastname: Dolinnaya
authors.forename: V
authors.initials: V
authors.lastname: Gajdos
authors.forename: O
authors.initials: O
authors.lastname: Burenina
authors.forename: E
authors.initials: E
authors.lastname: Fedotova
authors.forename: J S
authors.initials: JS
authors.lastname: Ellis
authors.forename: T
authors.initials: T
authors.lastname: Oretskaya
authors.forename: T
authors.initials: T
authors.lastname: Hianik
authors.forename: M
authors.initials: M
authors.lastname: Thompson
mesh_terms: D000162:Acoustics; D000818:Animals; D001483:Base Sequence; D004249:DNA Damage; D004563:Electrochemistry / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D017348:Microscopy, Electron, Scanning Transmission; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D013816:Thermodynamics; D013936:Thymidine / Q000031:analogs & derivatives*; D014466:Ultraviolet Rays
chemical_list: D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides; C029390:thymidine glycol; D013936:Thymidine