Arg-X Protease-Sensitive in Supramolecular Structures of Interphase Cell Nucleus during Growth Morphogenesis Mature Germs of Wheat

Abstract： In the present study was investigated Arg-X protease-sensitive in supramolecular-genome compartments （nucleoplasm, chromatin, nuclear matrix）, during the period of the transcriptional activation of chromatin when the growth processes was initiated in the mature germs of winter and transformed from it spring wheat. The germs have been separated from endosperm from 0 h （air-dry seed） up to 21 h in each 3 h after the start of seeds soaking. Cell nucleus have been allocated from germs and cleared, and then from them supramolecular-genome compartments were extracted by increasing ionic strength of solution. The Arg-X （tryptase） activity was assessed by cleavage of Arg-X bonds in the arginine-enriched protein protamine in all nuclear fractions. In the present study have shown what Arg-X protease-sensitives zones can be located on the supramolecular structures of chromatin matrix in processes of realization of ontogenetic programs of development in mature germs of the winter and transformed from it spring wheat. Arg-X protease-sensitive can translocate and coordinated in heteropolymer structures on the same genetic matrix. Questions of epigenetic mechanisms are discussed.

Arg-X Activity in Trypsin-like Complexes of the Nuclear Rroteins in the Suprastructures of Interphase Chromatin During Induction of Growth Morphogenesis Mature Germs of Wheat

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.