Changes in conformation, stability and condensation of DNA by univalent and divalent cations in methanol-water mixtures.
Circular dichroism spectroscopy, absorption spectroscopy, measurements of Tm values, sedimentation analysis and electron microscopy were used to study properties of calf thymus DNA in methanol-water mixtures as a function of monovalent cation (Na+ or Cs+) concentration and also in the presence of divalent cations Ca2+, Mg2+, and Mn2+. In the absence of divalent cations only slight conformational changes occurred and no condensation and/or aggregation could be detected. The Tm values depend on the amount of methanol and on the nature and concentration of cations. In methanol-water mixtures higher thermal stability was observed in solutions containing Cs+ ions. Up to 40% (v/v) methanol the addition of divalent ions leads to DNA stabilization. At methanol concentration higher than 50% the presence of divalent cations causes DNA condensation and denaturation even at room temperature. The denaturation is reversible with respect to EDTA addition indicating that no separation of complementary strands occurred and the resulting form of DNA is probably similar to the P form. DNA destacking appears to be a direct consequence of stronger cation binding by the condensed DNA in methanol-water mixtures.
Votavová
H
H
Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Sciences, Prague.
Kucerová
D
D
Felsberg
J
J
Sponar
J
J
D000818:Animals; D002412:Cations; D002413:Cations, Divalent; D002414:Cations, Monovalent; D002417:Cattle; D002942:Circular Dichroism; D004247:DNA / Q000648:ultrastructure; D006358:Hot Temperature; D000432:Methanol; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D013950:Thymus Gland; D014867:Water
D002412:Cations; D002413:Cations, Divalent; D002414:Cations, Monovalent; D014867:Water; D004247:DNA; D000432:Methanol