Left-handed DNA: intercalation of the cyanine thiazole orange and structural changes. A kinetic and thermodynamic approach.
The conditions under which different structures of left-handed DNA (poly(dG-me(5)dC)·poly(dG-me(5)dC)) can exist are investigated by spectrofluorometric, spectrophotometric, circular dichroism and calorimetric measurements and the kinetics of the transformations are analysed. The effects of temperature, salt and ethanol content on the transitions are also studied. The left-handed structure obtained by addition of either Mg(2+) ions or EtOH corresponds to Z-DNA, whereas the structure obtained using the mixture Mg(2+)/EtOH corresponds to the aggregate Z*-DNA. Upon addition of the fluorescent cyanine Thiazole Orange (TO), the transition Z → B immediately starts, whereas Z*-DNA retains its left-handed configuration in the presence of TO provided that the ratio [dye]/[polymer] ≤ 0.1. The equilibria and kinetics of the TO binding to Z*-DNA are investigated under the above conditions using the T-jump technique. The reaction mechanism consists of two series steps, the first one being characterized by the formation of an external electrostatic complex and the second corresponding to the dye penetration between the base pairs. A comparison with the B-DNA/TO system is drawn.
Biver
Tarita
T
Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Via Risorgimento 35, 56126 Pisa, Italy.
García
Begoña
B
Leal
José M
JM
Secco
Fernando
F
Turriani
Elisa
E
D052160:Benzothiazoles / Q000737:chemistry*; D002942:Circular Dichroism; D000431:Ethanol / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D007364:Intercalating Agents / Q000737:chemistry*; D007700:Kinetics; D009690:Nucleic Acid Conformation; D011089:Polydeoxyribonucleotides / Q000737:chemistry*; D011804:Quinolines / Q000737:chemistry*; D012492:Salts; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics
D052160:Benzothiazoles; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D011089:Polydeoxyribonucleotides; D011804:Quinolines; D012492:Salts; C051000:thiazole orange; D000431:Ethanol; C029988:poly(dG-m(5)dC).poly(dG-m(5)dC)