Biomedical subjects
Publications and source records attributed to Krylov DYu.
The conformational flexibility of DNA in transcriptionally active chromatin fractions has been estimated by circular dichroism spectroscopy analysis and was found to be restricted in the same fashion as in bulk chromatin. The observation is discussed in the context of different models of active chromatin organization.
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The size of DNA involved in the interaction with a histone octamer in H1-depleted chromatin was re-examined. We compared the thermal untwisting of chromatin DNA and naked DNA using CD and electrophoretic topoisomer analysis, and found that DNA of 175 +/- 10 base pairs (bp) in length interacted with the histone core under physiological conditions. The decrease of ionic strength below 20 mM NaCl reduced this length down to 145 bp: apparently, an extra 30 bp DNA dissociated from the histone core to yield well-known 145-bp core particle. Histone cores partly dissociate within the temperature range of 25 to 40 degrees C. Quantitative analysis of histone thermal dissociation from DNA shows that the size of DNA protected against thermal untwisting would be significantly overestimated if this effect is neglected. The results presented in this paper also suggest that the dimers (H2A, H2B) act as a lock, which prevents transmission of conformational alterations from a linker to nucleosome core DNA. The histone core dissociation as well as (H2A, H2B) dimer displacement are discussed in the light of their possible participation in the eukaryotic genome activation.
Relaxation of a DNA superhelical stress due to the B to A transition induced by trifluoroethanol has been studied by assessing the change of DNA orientation in a flow gradient. Using DNAs of different superhelical densities, a decrease in the winding angle during the B----A shift of DNA was found to be 1.5 degrees per base pair in solution. Accepting the winding angle for B-DNA in solution to be 34.1 degrees, that for A-DNA must have a value of 32.6 degrees which agrees with the X-ray data for A-DNA in the condensed state. The date obtained within the B-A transition interval make it possible to conclude that there is an increase in winding at each B/A junction, which is about 5 degrees per one junction.
Experimental phase diagrams (A form, B form, Coil) were built in the coordinates (a, alcohol fraction: T, temperature) for the natural DNAs and poly d(A-T). The main parameter of the B-A transition,-cooperativity length, nu o, was estimated by the slopes of the branches A-B, A-Coil, B-Coil in the vicinity of the triple point: nu o = 10-20 base pairs, which corresponds to the energy for the B/A junction of 1.2-1.8 kcal/mol. We discovered two new effects which are due to the coexistence of the three different conformations in one polymeric molecule: an increase in the melting temperature above that for the 'ideal' triple point (i.e. for the case of the ideal phase transitions); a widening of the melting curve within the B-A transition range.
The B-A transition is characterized by two main physical parameters which might be biologically important: the cooperativity length and free energy difference between the B and A states under physiological conditions. Earlier these values were determined by us in an experiment over the B-A shift in water-non-electrolyte solutions in the presence of small molecules ("ties") affecting the B-A equilibrium. Now we report a new method of determining the cooperativity length which utilizes a phase diagram (B,A, coil). The coordinates are the fraction of non-electrolytes and temperature. Application of the Ising model for joint description of the B-A and helix-coil transitions makes possible to find the cooperativity length using the known thermodynamic parameters of the DNA melting and the appearance of the phase diagram near the triple point (A,B,coil). The value thus found (approximately 10(1) base pairs) is in accord with the values obtained with ties. In the new method the junctions between the A and B segments actually play the role of ties, stabilizing the double-stranded state. A considerable effect of the melting curve widening within the B-A transition range was discovered. A possible explanation suggests the presence of the A-philic sequences in the natural DNA. The A-phility of the oligo G oligo C sequence was estimated from the B-A transition curves of the synthetic decanucleotide duplexes.