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An increase in the activity of histone-associated rat thymus nucleus proteinases specific for histones H2A, H2B and H1 was shown after gamma irradiation or hydrocortisone treatment of animals. Histone H1-specific proteinase activity is dependent on DNA and increases in the presence of denatured DNA, whereas proteinases specific for core histones are inhibited in the presence of denatured DNA. The increase in the activity of histone-associated proteinases depends on the radiation dose and the time after irradiation or hydrocortisone injection. In the presence of dithiotreitol and sodium dodecyl sulfate, these proteinases dissociate from histones. It was found by gel electrophoresis that several proteinases of various molecular masses are closely associated with histones obtained from thymus nuclei of irradiated or hydrocortisone-treated rats.

We investigated the effect of various forms of DNA (double- and single-stranded calf thymus DNA, circular plasmid DNA, gamma- and UV-irradiated DNA and DNAase I-treated double-stranded DNA) aggregated with histones, on the proteolysis of these histones by proteinase associated with the rat liver nuclear scaffold. It was shown that the nuclear scaffold-associated proteinase is able to degrade selectively the histone H1 only in the presence of the DNA containing single-strand breaks induced by gamma-radiation or DNAase I treatment as well as in the presence of heat-denatured DNA. This proteinase is not activated by the double-stranded circular plasmid DNA or by UV-treated double-stranded DNA. Histone H1-specific proteinase (HSP) activated by gamma-irradiated DNA is inhibited by inhibitors of serine proteinases such as antipain, leupeptin, phenylmethylsulphonyl fluoride, as well as by dithiothreitol. The results lead us to suggest that DNA-activated HSP from rat liver nuclei is involved in the regulation of the access of repair enzymes to the damage portions of DNA within chromatin.

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Acid-soluble proteins able to form DNA-protein complexes in the presence of physiological concentration of NaCl were isolated from rat liver mitochondria. Electrophoretic analysis of these proteins in 15% polyacrylamide gel showed that mitochondrial acid-soluble proteins include of approximately 20 polypeptides with molecular weight of 10-120 kDa. The fraction of acid-soluble proteins can be separated into basic and acidic proteins by chromatography on DEAE cellulose. Some of acidic proteins are tightly bound to the basic proteins and can be separated from them in the presence of 5 mM dithiothreitol. It is discovered that the fraction of acidic proteins contains proteases (including DNA-activated ones), which cleave different polypeptides of the basic proteins with different efficiency. Possibly, mitochondrial DNA-binding proteins and DNA-activated proteases are involved in the regulation of structural organization and functional activity of mitochondrial DNA.

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