The purpose of this study was to analyze spatio-temporal dynamics of localization of protease-sensitive sites Arg-X in non-histone and histone blocks of heteropolymer suprastructures （nucleoplasm, chromatin, nuclear ...The purpose of this study was to analyze spatio-temporal dynamics of localization of protease-sensitive sites Arg-X in non-histone and histone blocks of heteropolymer suprastructures （nucleoplasm, chromatin, nuclear matrix） as possible zones affecting the conformational rearrangements of the total interphase chromatin at the induction of increasing morphogenesis of mature embryos-germs of spring and transformed from its winter wheat. Germinated embryos-germs were detached from endosperm after 24 hours from the start of soaking. Cell nuclei have been allocated from embryos-germs and cleared, and then from their heteropolymer suprastructures （nucleoplasm, chromatin loosely bound with nuclear matrix and chromatin tightly bound with nuclear matrix, and nuclear matrix） were extracted by increasing ionic strength of solution. From isolated nuclear suprastructures, non-histone proteins were separated from histones using ion exchange chromatography. Trypsin-like complexes from non-histone proteins and histone blocks were isolated using the affinity chromatography. The Arg-X （tryptase） activity was assessed by cleavage of Arg-X bonds in the arginine-enriched protein protamine. Hypersensitivity to the Arg-X proteolysis in trypsin-like complexes detected at the level suprastructures of chromatin tightly bound with the nuclear matrix was shown. The most active changes of the nuclear proteome have occurred at the level of the non-histone proteins and the core histones （H2A ＋ H2B） （H3 ＋ H4） of induced to growth embryos-seedlings of winter wheat （compared to the initial spring form of wheat）. Perhaps hypersensitivity to the Arg-X activity of the trypsin-like complexes in the non-histone proteins and the core blocks of chromatin tightly bound with nuclear matrix have been entrenched during the transforming of the winter wheat from the initial spring wheat.展开更多
文摘The purpose of this study was to analyze spatio-temporal dynamics of localization of protease-sensitive sites Arg-X in non-histone and histone blocks of heteropolymer suprastructures （nucleoplasm, chromatin, nuclear matrix） as possible zones affecting the conformational rearrangements of the total interphase chromatin at the induction of increasing morphogenesis of mature embryos-germs of spring and transformed from its winter wheat. Germinated embryos-germs were detached from endosperm after 24 hours from the start of soaking. Cell nuclei have been allocated from embryos-germs and cleared, and then from their heteropolymer suprastructures （nucleoplasm, chromatin loosely bound with nuclear matrix and chromatin tightly bound with nuclear matrix, and nuclear matrix） were extracted by increasing ionic strength of solution. From isolated nuclear suprastructures, non-histone proteins were separated from histones using ion exchange chromatography. Trypsin-like complexes from non-histone proteins and histone blocks were isolated using the affinity chromatography. The Arg-X （tryptase） activity was assessed by cleavage of Arg-X bonds in the arginine-enriched protein protamine. Hypersensitivity to the Arg-X proteolysis in trypsin-like complexes detected at the level suprastructures of chromatin tightly bound with the nuclear matrix was shown. The most active changes of the nuclear proteome have occurred at the level of the non-histone proteins and the core histones （H2A ＋ H2B） （H3 ＋ H4） of induced to growth embryos-seedlings of winter wheat （compared to the initial spring form of wheat）. Perhaps hypersensitivity to the Arg-X activity of the trypsin-like complexes in the non-histone proteins and the core blocks of chromatin tightly bound with nuclear matrix have been entrenched during the transforming of the winter wheat from the initial spring wheat.
