Antigenic properties of heterogeneous nuclear ribonucleoprotein particles weakly binding nonhistone proteins and proteins binding single-stranded DNA (SSB proteins) from Ehrlich ascites tumor]

Antigenic properties of the proteins of heterogeneous nuclear ribonucleoprotein particles, (hnRNP), weakly bound nonhistone chromatin proteins (WB(N)P) and single-strand DNA-binding proteins (SSB proteins) from chromatin and extrachromatin fraction of the Ehrlich ascites tumor cells have been comparatively studied. The chromatin and extrachromatin SSB proteins displayed similar mobility in the tube and slab SDS/PAGE, had the same ssDNA-binding capacity and similarly stimulated the replicative synthesis in permeable cells. However, the chromatin SSB proteins contained 1.4 times higher phosphate amount than the extrachromatin ones (3.1 and 2. 2. moles phosphorus per 1 mole protein, respectively). The study of four protein groups with the use of a rabbit antiserum to/against extrachromatin SSB proteins (titer 1:13000 by enzyme immunoassay) showed that the chromatin and the extrachromatin SSB proteins have similar antigenic properties. One fraction of the hnRNP proteins was also reactive with the antiserum, whereas the WB(N)P displayed no cross-reactivity. The specificity of the ferm "SSB proteins" as applied to eukaryotic cells, their affinity with hnRNP proteins and differences from the HMG proteins are discussed.     

The microelectrophoresis of the DNA in individual intact and gamma-irradiated thymocytes

The in vitro gamma-irradiated mouse thymocytes were embedded in low melting agarose at 37 degrees C. After getting at 4 degrees C, the cells were lysed in neutral detergent solution containing proteinase K and ethidium bromide. Microscopic visualization of single lysed and stained cells showed the presence of the central "core" (nuclear matrix) surrounded with "halo" (relaxed nuclear DNA). During electrophoresis (2-5 V/sm, 5 min) this "halo" migrated towards the anode forming a "tail". The use of microdensitometric system provided measuring the size of the tail (L) and quantity of migrated DNA (S) for individual cells as well as obtaining the distribution of these parameters among the cells. The latter may be characteristic of heterogeneity of the cell population. It was shown that L and S increased linearly with the dose irradiation at least between 0.2 and and 5.0 Gy. In irradiated thymocyte (3 Gy) the DNA repair occurred within 10-20 min, but residual DNA damage could be observed even after 60 min of incubation. These damages may initiate the degradation of DNA in irradiated thymocytes that was observed after the repair of DNA.     

Participation of the SSB protein in the repair of the DNA of Ehrlich ascites carcinoma cells following UV irradiation

Synchronous changes were detected in the SSB-protein content of the chromatin and in the rate of repair DNA synthesis at different time intervals after UV-irradiation of Ehrlich ascites tumor cells. The amount of SSB-protein in the extra-chromatin fraction was in an inverse relation to its content in the chromatin, whereas the cumulative SSB-protein content remained invariable. Similar changes in the SSB-protein content of the chromatin and in repair synthesis were also registered after the effect of various doses of UV-light. The increase of the SSB-protein content in the chromatin was not connected with the postirradiation accumulation of single-strand sites in DNA.     

Role of hypoxanthine in the radiation death of thymocytes in vitro

Hypoxanthine in vitro causes death of thymocytes and concurrent intranucleosome degradation of chromatin. This process is more manifest in a more radiosensitive thymocyte fraction and prevented by protein synthesis inhibitors. The increase in the yield of hypoxanthine after the effect of lympholytic agents of different nature is not the result of cell death. It is assumed that hypoxanthine, formed in the exposed cells, may be an additional cytotoxic factor on reaching a subliminal concentration.     

Nature of the death of thymus lymphocytes as affected by different kinds of harmful factors

It was shown that thymus lymphocyte death caused by the effect of dithiobisdinitrobenzoic acid, hyperthermia, and osmotic shock is not accompanied by ordered chromatin degradation. Protein synthesis inhibitors do not prevent thymocyte death under these effects. The authors discuss the kinds of lymphoid cell death which depend upon the nature of the damaging factor and differ both in morphology and the character of nuclear material degradation.     

Spectrophometric analysis of volatile compounds in microorganisms

A simple modification of a spectrophometric method was proposed for the rapid detection of microorganisms based on their ability either to excrete or to absorb volatile compounds. The method provides the possibility of contactless control for bacterial growth at a concentration above 10⁷ cells/ml. In addition, the method allows discriminating mutants of the fungus Neurospora crassa defective in the nitrogen metabolism from the wild type strains. It is likely that nitrite reductase and nitrate reductase enzymes regulated by the nit-2 and nit-6 genes are involved in formation of the water soluble volatile compounds of this organism.