The role of spontaneous DNA damage in the radiosensitivity of thymus lymphocytes

The most radiosensitive thymus cortical lymphocytes are highly sensitive to inhibitors of repair of spontaneous DNA lesions known by endonucleases: this is indicative of different level of injury to cells. The preincubation of cells with chemical inductors of differentiation increases the number of spontaneous lesions in them. The preincubation of thymocytes with concanavalin A removes almost completely the differences in the level of injury.     

Specific activity and molecular forms of NAD-kinase in Neurospora crassa ontogeny

The specific activity and molecular forms of NAD-kinase during ontogenesis of Neurospora crassa were investigated. The specific activity of the enzyme increased drastically at critical stages of fungal development, i.e. during conidia germination and during transition from the logarithmic to stationary growth stage, reaching 85 nmole NADP/hr/mg protein. By polyacrylamide gel electrophoresis four forms of NAD-kinase were revealed that had the following molecular masses: I-338,000, II-306,000, III-229,000, and IV-203,000. The vegetative mycelium contained predominantly form III, and conidia showed a high content of high-molecular-weight forms I and II.     

Effect of the synthetic Leu enkephalin dalargin on the protein and nucleic acid content of the muscles in the rainbow trout

After dalargin treatment of fish eggs (at the stage of swollen egg envelopes) and on juveniles (at the stage of early meiotic oocyte appearance in gonads) DNA content is observed to raise up to 85% and 66%, respectively in yearlings and up to 20% and 23% respectively in second-year fish as compared to the control ones. Dalargin also influences the exchange of muscle RNA in yearlings and second-year rainbow-trout. Dalargin effect is higher when peptide influences at the very onset of organism differentiation.     

Molecular mechanisms of radiation death of lymphoid cells. Radioprotective effect of cysteamine on a subpopulation of thymocytes differing in radiation sensitivity

The radioprotective effect of cysteamine, incubated with thymocytes for 20 min prior to gamma-irradiation, was much more manifest with regard to subpopulations of radiosensitive small cortical cells (DMF = 1.8) than large thymocytes (DMF = 1.2). Radioprotective concentrations of cysteamine relaxed completely superhelical DNA of thymocytes. The relaxation was not due to the formation of single-strand breaks. A partial recovery of superhelical conformation of DNA of small thymocytes before irradiation reduced the radioprotective effect of cysteamine.     

Stimulation of replicative DNA synthesis by eukaryotic proteins bound to single-stranded DNA]

The paper deals with the effect of the single-strand (ss) DNA-binding proteins (SSB-proteins) from the Ehrlich ascites tumor (EAT) cells and from the eggs of silkworm, as well as the mouse serum blood proteins, having preferential affinity to ss DNA, on the DNA replicative synthesis in the EAT cells permeable for the macromolecules, and, for the silkworm proteins and on the DNA replicative synthesis in the nuclei from the eggs of silkworm proteins and on the DNA replicative synthesis in the nuclei from the eggs of silkworm permeable for macromolecules. SSB-proteins of EAT to considerable extent stimulated the DNA synthesis. At the same time, the other proteins (from the silkworm and from the serum) activated the DNA synthesis in the permeable cells to the less extent. It was found that SSB-proteins from the silkworm had a 1.5-13 fold stimulating effect on the DNA replicative synthesis in the homologous system (in the permeable nuclei). If the permeability for the macromolecules of the cells and nuclei treatment with Triton X-100 may be different, it is supposed that the activation of the DNA synthesis by the exogenous proteins depends on the homologous system of the DNA replicative complex. It is possible that the effect of the serum proteins on the DNA synthesis is connected with the masking of the ss regions of DNA which inhibited DNA-polymerase alpha. Perhaps the mechanisms of the activation of the DNA replicative synthesis by the proteins in vitro with the purified DNA polymerase alpha and in vivo are of different nature and are conditioned by homology of the deoxyribonucleoproteins.     

The effect of SSB-protein from Ehrlich ascites carcinoma cells on activity of various enzymes of DNA replication and repair

The effect of a single-stranded DNA-binding protein (SSB-protein) form Ehrlich ascites tumour cells (EAT) on the activity of homologous purified DNA-polymerases alpha and beta, DNA-replicase, primase and DNA-polymerases from phage T4 and Bacillus stearothermophillus was studied. It was shown that the SSB-protein caused a 1.5-2.5-fold stimulation of the DNA-polymerase alpha activity on different templates (e.g., denaturated and activated DNA, poly(dA). The degree of stimulation depended on the template type, protein/template ratio and purity of DNA-polymerase alpha. The activity of DNA-polymerase was inhibited by the SSB-protein, when the activated DNA was used as a matrix and was unchanged on the denaturated DNA. The activity of some prokaryotic DNA-polymerases was increased under the influence of the SSB-protein. The protein enhanced the processivity of T4 DNA-polymerase and strongly inhibited the activity of replicase and primase. A conclusion about the complex effect of the SSB-protein on the activity of replicative and repair enzymes is drawn.     

Bacterial NO synthases

Unlike mammalian NO synthases, bacterial NO synthases do not contain a reductase domain. The only exception from this rule is the NO synthase from myxobacterium Sorangium cellulosum, but its reductase domain has unusual structure and location in the enzyme molecule. Recent achievements in bacterial genome sequencing have revealed the gene coding NO synthase (represented as an oxygenase domain) in some bacteria and have advanced the study of structure and functions of bacterial NO synthases. Important features of structure, sources of reducing equivalents, evolutionary connections, and functions of bacterial NO synthases (i.e. participation in nitration of the indole ring of Trp, in reparation of UV-radiation damage, role in adaptation of bacteria to oxidative stress, participation in the synthesis of cGMP, and resistance of bacteria against antibiotics) are described.     

Poly(ADP-ribosylation) of proteins determines the pool of endogenous NAD and the radiosensitivity of thymus lymphocytes

A decrease in the endogenous NAD content that occurs immediately after gamma-irradiation of thymus lymphocytes is attributed to activation of poly (ADP-ribosylation) and not to changes in the activity of NAD-glycohydrolase and/or to the release of NAD from cells. The addition of benzamide 60 min before irradiation prevents the postirradiation drop of the NAD level and produces a radioprotective effect. At the same time, benzamide inhibits nuclear superhelix DNA repair and causes an average of 40 per cent decrease in the activity of DNA ligases I and II. The authors discuss the idea that the content of intracellular NAD in thymocytes is a critical factor responsible for the vitality of these cells.