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Immunosuppressive factor (ISFnp) which inhibits proliferation and viability of thymoma EL-4 cells was isolated from mouse liver. The testing procedure based on the biotransformation of MTT-tetrazolium by mitochondrial enzymes of viable cells allowed us to purify this factor as individual peak of protein, that allowed to obtain polyclonal rabbit antibodies to this factor. By the methods of double immunodiffusion, gel-filtration and SDS-PAGE with subsequent immunoblotting we shown that this factor specifically localized in liver and consists two subunits of 40 and 42 kDa which form dimers with apparent M(r) about 70-80 kDa. This factor induced olygonucleosomal DNA cleavage in EL-4 cells in vitro similar to dexamethasone-induced DNA-degradation in thymocytes. This cleavage preceded to lysis of EL-4 cells assessed by 51Cr-release, that strongly suggested an involvement of apoptosis in cell death mechanism. ISFnp strongly inhibited blast-transformation and proliferation in MLC-responses to mutant MHC class 2 molecule. This effect was not due to deletion of allo-reactive clones, because removing of this factor from MLC cultures treated with one for 4 days resulted in blast-transformation without any reduction of the number of viable cells as well as their capacity for secondary responses to the same antigen as compared with control cultures.

The rate constants of 1H--3H exchange between water and C8H-groups of purinic residues of DNA in solution and in nuclei of synchronous Physarum polycephalum at different phases of the cell cycle were measured. The nuclear membranes and nonhistone proteins were shown not to create additional hindrances and not to diminish the availability of DNA in nuclei for solvent molecules. A small increase of the rate of the 1H--3H exchange between water and the DNA of nuclei upon transition from the S-phase to the late G2-phase seems to reflect the process of chromatin decondensation connected with activation of the transcription and local changes of the secondary structure of DNA at the late G2-phase of the cell cycle.

The rate constants of 1H----3H exchange between water and C8H-groups of purine residues of alternating polynucleotides: poly[d(A-C)].poly[d(G-T)] and poly[d(A-T)].poly[d(A-T)], as well as Escherichia coli DNA, dAMP and dGMP, in solutions with high concentration (4.3 or 6 M) CsF, in water ethanol (60%) solution and (in comparison) in 0.15 M NaCl were determined at 25 degrees C. The 1H----3H exchange rate exchange rate constants for adenylic (kA) and guanylic (kG) residues of polynucleotides were compared with the corresponding constant for DNA and mononucleotides. It was shown that at conditions when poly[d(G-T)] and poly[d(A-T)].poly[d(A-T)] exhibit the "X-form" CD spectrum, alteration of exchange rates in polynucleotides (approximately 2-fold increase in kA in CSF and approximately 1.5-fold decrease in kA and kG in 60% ethanol with 0.15 M NaCl) is due to the effect of solvents on the chemical reactivity of purine residues, but does not reflect a conformational transition. The analysis of these results allows us to conclude, that alternating polynucleotides under the above mentioned conditions retain roughly the conformations inherent in them in 0.15 M NaCl: poly[d(A-C)].poly[d(G-T)] conformation in 4.3 m CsF or 60% ethanol differs only insignificantly from the "canonic" B-DNA, whereas the poly[d(A-T)].poly[d(A-T)] conformation in 6 M CSF corresponds to B-alternating DNA.

The rate constants of 1H----3H exchange between water and C8H-groups of purinic residues of alternating polynucleotides: poly[d(A-T)].poly[d(A-T)] (I), poly[d(G-C)].poly[d(G-C)] (II), poly[d(A-C)].poly[d(G-T)] (III) and homopolynucleotides: poly(dA).poly(dt) (IV), poly(dG).poly(dC) (V), as well as DNA E. coli, was determined in 0.15 M NaCl at 25 degrees C. The retardation of exchange observed at these conditions (compared to that of the B-form DNA) is in agreement with the model of B-alternating structure for the (I) and is attributed to the co-existence of B- and A-conformers for the (V) in solution. Absence of distinguishable differences in exchange rate constants for purinic residues of the (II), (III) and (IV) (compared to that of the B-form DNA) evidences that conformations of these polynucleotides in solution are similar to "canonical" B-form DNA and don't correlate with the model of "heteronomous" DNA which was proposed for (IV).