Mg2+ and Ni2+ ion effect on stability and structure of triple poly I.poly A.poly I helix.

The effects of Mg2+ and Ni2+ ions on the absorption spectra of IMP, single-stranded poly I and three-stranded A2I in solutions with 0.1 M Na+ (pH 7) have been studied. In contrast to Mg2+ ions, the Ni2+ ions affect the absorption spectra of these polynucleotides and IMP. The concentration dependences of the intensity at the extrema in the differential UV spectra suggest that in the region of high Ni2+ concentrations ionic complexes with poly I and A2I are formed, which are characterized by the association constants K'''I = 2000 M(-1) and K'''A2I = 550 M(-1), respectively. The shape of the DUV spectra prompts the conclusion that these complexes are formed due to the inner-sphere interaction of Ni2+ ions with N7 of poly I and A2I presumably due to the outer-sphere Ni2+-O6 interaction. The formation of the complexes leads to destruction of A2I triplexes. The dependences of the melting temperature (T(m)) of A2I on Mg2+ and Ni2+ concentrations have been measured. The thermal stability is observed to increase at the ionic contents up to 0.01 M Mg2+ and only to 2x10(-4) M Ni2+. At higher contents of Ni2+ ions, T(m) lowers and the cooperativity of A2I melting decreases continuously. In all the cases the melting process is the A2I-->A+I+I (3-->1) transition. According to the "ligand" theory, these effects are generated by the energy-advantageous Ni2+ binding to single-stranded poly I (K'''A2I < K'''I) and by the greater number of binding sites which appears during the 3-->1 transition and is entropy-advantageous.

[Interaction of divalent metal ions with four-stranded polyriboinosinic acid]. / Vzaimodeistvie ionov dvukhvalentnykh metallov s chetyrekhtsepochechnoi poliriboinozinovoi kislotoi.

Interaction of Mg2+, Ca2+, Cu2+ ions with the four-stranded poly(I) was studied using differential UV and visible spectroscopies. It was shown that, up to concentrations of approximately 0.1 M, Mg2+ and Ca2+ ions do not bind to heteroatoms of hypoxanthine of the four-stranded poly(I). Cu2+ ions interact with N7 (and/or N1) and O6 (through the water molecule of the hydrate shell of the ion). The latter seems to induce the enolization of hypoxanthine the deprotonation of N1, and, as a result, the transition of the four-stranded helix to single-stranded coils. Single-stranded chains form compact particles with an effective radius of about 100 A.

[Interaction of Ag+ ions with DNA and its monomers]. / Vzaimodeistvie ionov Ag+ s DNK i ee monomerami.

Differential UV spectra of DNA and its monomers that were induced by Ag+ ions were measured, and the effect of ions on the parameters of the helix-coil transition was studied. The data obtained confirm the existence of "strong" and "weak" modes of binding of Ag+ to DNA. The earlier proposed proton transfer from N1G to N3C, which is determined by the interaction of Ag+ with N7G (a "strong" complex), follows immediately from the shape of the differential UV spectra. The positive cooperativity of the binding of Ag+ to DNA upon the formation of a "weak" complex is due to the cooperativity of the transition of DNA to a new double-helical conformation. A model of this conformation is proposed which suggests the formation of Hougsteen GC and AT pairs.

[Interaction of Ag+ ions with ribonucleotides of canonical bases]. / Vzaimodeistvie ionov Ag+ s ribonukleotidami kanonicheskikh osnovanii.

The interaction of Ag+ ions with ribonucleotides of canonical bases in aqueous solution was studied by differential UV spectroscopy. Atoms coordinating silver ions (N7, O6 of guanosine 5'-monophosphate, N3, O2 of cytidine 5'-monophosphate, N7, N1, N3 of adenosine 5'-monophosphate and N3 of uridine 5'-monophosphate) and the binding constants characterizing the formation of appropriate complexes were determined. The differences in the relative affinity of Ag+ ions for the atoms of nucleotide bases correlate with the potential on them.

[Interaction of divalent cadmium ions with nucleotides and native DNA]. / Izuchenie vzaimodeistviia ionov dvukhvalentnogo kadmiia s nukleotidami i prirodnoi DNK.

Complex formation of Cd2+ ions with 2'-deoxy-5'-phosphates of canonical bases and their riboanalogues in water solution is studied by the method of differential UV-spectroscopy. It is stated that the atoms coordinating Cd2+ in complexes are N7 of dGMP and GMP, dAMP and AMP, N1 of adenine derivatives, N3 of dCMP. No interaction with base heteroatoms of UMP and dTMP is found. O2' present in the structure of the sugar ring has a weak influence on the Cd2+ ion binding to purine nucleotides. It manifests itself strongly in the complexes with cytosine derivatives: cadmium does not interact with N3 of CMP and poly-C practically. In the last case O2 is a centre coordinating Cd2+ ions. The interaction with this atom induces the melting of polynucleotide helical parts. At the high cadmium concentration poly-C forms compact particles. The main centre binding Cd2+ ions in DNA is N7 of guanines. Noncooperative interaction with these centres results in the internal protonation of N3C of GC-pairs. This is not followed with the disordering of the DNA helical structure.

[The "condensation" and "ligand" theory in describing DNA helix-coil transition: comparative analysis]. / Teorii "kondensatsii" i "skrepok" pri opisanii perekhoda spiral'-klubok DNK: sravnitel'nyi analiz.

Experimental data on DNA melting in the presence of magnesium and calcium ions are compared with the calculated results by the "ligand" theory and "condensation" theory developed by de Marky and Manning (TMM). The maximum is observed in TMM dependences of DNA melting temperature change (delta Tm) on the ion concentration (D), absent in the experimental dependences and the "ligand" theory. The main cause of the discrepancy of calculated and experimental dependences delta Tm (D) is the fact that TMM neglects Na+ ion effects on Me2+ ion binding ("condensation") to DNA. The results obtained show that in comparison with TMM the "ligand" theory describes more adequately DNA helix-coil transition in the presence of divalent metal ions in the region of both low and high ionic strengths of solution.

Interaction of bivalent copper, nickel, manganese ions with native DNA and its monomers.

UV differential spectroscopy is applied to study the interaction of Cu2+, Ni2+, Mn2+ ions with deoxyribonucleotides of canonic bases (dGMP, dAMP, dCMP, dTMP) and native DNA. Heteroatoms of the bases, coordinating ions, and binding constants which characterize the formation of metal complexes are found. The affinity of the ions is lower for the deoxyribonucleotide bases than for the ribonucleotide ones. This indicates that 02' of ribose participates in the stabilization of the metal complex even under conditions close to the neutral one (pH 6). Unlike the Cu2+ ions, Ni2+ and Mn2+ ions do not interact with N3C both in monomers and polymers. This seems to be the main factor explaining why copper makes DNA transform into a structure with a quasi-Hoogsteen pairing of GC pairs. No transformations of this kind of helix-coil transitions are caused by manganese and nickel up to concentrations 4 X 10(-2) M.

[Structural and physicochemical characteristics of DNA from animal tissue subjected to chronic irradiation in the Chernobyl area]. / Strukturnye i fiziko-khimicheskie kharakteristiki DNK iz tkanei zhivotnykh, podvergavshikhsia dlitel'nomu khronicheskomu oblucheniiu v zone chernobylia.

The properties of animal DNAs exposed to prolonged irradiation in the Chernobyl zone, have been studied by the methods of viscometry, thermal denaturation, IR-spectroscopy, and electrophoresis. High content of low-molecular fractions have been observed in the preparations of DNA from liver and spleen, their quantities increasing with age and generation of animals. This effect is especially strong in DNA from liver. Low-molecular fraction of DNA is shown to be enriched with G-C pairs and to consist of the following four fractions: 1) approximately 500 base pairs (B.p.), 2) approximately 1.5 divided by 2 thousand B.p., 3) approximately 4 divided by 5 thousand B.p. and 4) a mixture of approximately 20 thousand B.p. fragments. Further, it has been observed that the DNA preparations from the tissues of experimental animals contain about ten times higher contents of Fe, Zn, Se and other elements as compared to the control preparations.

[The study of complex-formation of DNA with the antimicrobial drug decamethoxine]. / Izuchenie kompleksoobrazovaniia DNK s antimikrobnym preparatom dekametoksinom.

The interaction of effective antibacterial drug decametoxyn with natural DNA was studied by UV-spectroscopy. Decametoxyn shows a specificity to nucleotides: it decreases the cooperativity of melting and the thermal stability of DNA parts enriched by AT pairs. The characteristics of the helix-coil transition on the DNA parts enriched by GC-pairs are invariable. Interaction with AT-pairs results in their partial or complete melting at room temperature, followed by intermolecule aggregation. Interacting with phosphates decametoxyn manifests itself not as a dication but as two single-charged ions.

Studies of formation of bivalent copper complexes with native and denatured DNA.

The formation of Cu2+ complexes with native and denatured DNA is studied by the methods of differential UV spectroscopy, CD spectroscopy, and viscometry. On ion binding to the bases of native DNA the latter transforms into a new conformation. This transition is accompanied with a sharp increase in UV absorption and a decrease in the intrinsic viscosity though the high degree of helicity persists. Possible sites of Cu2+ ion binding on DNA of various conformations are found along with corresponding constants of complex formation.

[Binding of ions of trivalent iron with DNA]. / Issledovanie sviazyvaniia ionov trekhvalentnogo zheleza s DNK.

The DNA helix-coil transition in nonbuffer solutions of Fe(NO3)3 was studied. Calculation of the ionic equilibrium indicated that in these solutions iron exists in the form of mono-, bi- or trivalent hydroxide, the formation of which decreases pH. A component of the DNA thermal stability variation associated with DNA binding to iron ions was calculated. An increase in the iron contents produces an increase in the melting range which was determined by a rise in the melting end temperature when binding the ions with phosphates and a drop in the melting beginning temperature when binding to DNA bases. A main contribution to the former effect is made by [Fe2(OH)3]3+ ions and to the latter effect by [FeOH]2+ ions. The constants of ion binding are higher for bases than for phosphates. Differential UV spectra of native and denatured DNA due to iron ions were measured. Calculations of conformation and coordination components of these spectra show that G-C pairs are one of the possible sites of iron ion binding with DNA.

[Effect of divalent copper ions on heat denaturation of DNA]. / Issledovanie deistviia ionov dvukhvalentnoi medi na teplovuiu denaturatsiiu DNK.

Using the thermal denaturation method the effect of bivalent copper of (4-10(-6)-10(-3)) M concentrations on the helix-coil transition of DNA was studied in the solution of Na+ concentrations 10(-3)-10(-1) M. Unlike the previous studies, this paper makes allowance for the effect of impurity ions present in DNA and deionized water. It has been shown that in the region of low Cu2+ and Na+ concentrations, thermal stability increases, the melting range extends and the denaturation curves become asymmetric. At concentrations more than approximately 3-10(-5) M Cu2+, melting temperature starts to fall, and the range reduces to 1-1.5 degrees at [Cu2+] greater than or equal to 2-10(-4) M. As [Cu2+] reaches these values, the denaturation curve asymmetry and melting range increase again, which is due to the inversion of the relative stability of AT- and GC-pairs. Employing experimental and phase-transition-theory data for homopolymers, the constants of Cu2+ binding with phosphates and DNA bases were calculated. The concentration dependence of the DNA denaturation parameters was shown to be governed by the superposition of binding Cu2+ with phosphates and nucleic acid bases.

[Effect of magnesium ions on heat denaturation of DNA]. / Issledovanie deistviia ionov magniia na teplovuiu denaturatsiiu DNK.

The dependence of animal DNA denaturation on magnesium ion concentration has been studied in the range (10(-6)--10(-1) M with sodium ion content of 10(-3) and 10(-2) M. Special attention has been given to the effect of multivalent metallic impurities bound to DNA. An increase of DNA thermal stability has been shown to occur in the magnesium concentration rage of 10(-6)--10(-4) M. At concentrations exceeding 10(-3) M the T M begins to decrease. The dependence of the DNA melting range on magnesium ion concentration has a maximum at approximately 10(-5) M Mg2+. At low magnesium and sodium ion concentrations a strong asymmetry of the melting curves has been observed. This effect can be described in terms of the melting theory for DNA complexed with small molecules and is explained by magnesium ion redistribution from the denatured portions of DNA to native ones. The method for calculation of melting curves in the DNA-ligand system has been proposed. Studies of thermal denaturation parameters have been shown to be an effective method for the estimation of binding constants of ligands to native and denatured DNA.