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首页> 美国卫生研究院文献>Nucleic Acids Research >The non-histone proteins of the rat liver nucleus and their distribution amongst chromatin fractions as produced by nuclease digestion.
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The non-histone proteins of the rat liver nucleus and their distribution amongst chromatin fractions as produced by nuclease digestion.

机译：大鼠肝核的非组蛋白蛋白及其在核酸酶消化产生的染色质组分中的分布。

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摘要

The search for proteins involved in maintaining higher order chromatin structures has led to a systematic examination of the non-histone proteins (NHP) of rat liver nuclei in the context of nuclease digestion studies. 40-45% of the 3H-tryptophan labelled NHP originally present could be removed by extensive washing in a "physiological" buffer, incubation at 37 degrees C with or without nuclease and a further wash step. Nuclei at this stage had a remarkably constant NHP content (ca. 0.73 micrograms/micrograms DNA), independent of the degree of digestion with micrococcal nuclease or HaeIII. The solubilized chromatin produced by limited digestion with either nuclease contained 0.3-0.5 microgram NHP/microgram DNA, this value falling to ca. 0.16 after more extensive cleavage. Insoluble chromatin fractions were between 2-fold (very limited digestion) and 16-fold (extensive digestion) richer in NHP than the corresponding soluble fractions. Gel electrophoresis revealed about 12 NHP bands in soluble fractions, the most prominent of M.Wt. 41.400, while the insoluble material had at least 50 components. These properties were independent of whether lysis of nuclei occurred in 0.2 or 50 mM ionic strength. The large disparity in NHP content between complementary soluble and insoluble chromatin fractions is considered in terms of chromatin organization in vivo and the possible role of NHP migration.
机译：对涉及维持高级染色质结构的蛋白质的搜索已导致在核酸酶消化研究的背景下系统检查大鼠肝核的非组蛋白。最初存在的3H-色氨酸标记的NHP的40-45％可以通过在“生理”缓冲液中充分洗涤，在37摄氏度下（无论有无核酸酶）孵育和进一步的洗涤步骤除去。在此阶段，核的NHP含量非常恒定（约0.73微克/微克DNA），与微球菌核酸酶或HaeIII的消化程度无关。通过用任一种核酸酶有限消化而产生的增溶的染色质包含0.3-0.5微克NHP /微克DNA，该值降至约10。更广泛的卵裂后为0.16。不溶性染色质组分的NHP含量比相应的可溶组分高2倍（非常有限的消化）至16倍（广泛消化）。凝胶电泳显示可溶级分中约有12条NHP条带，是M.Wt中最突出的条带。 41.400，而不溶物质至少具有50种成分。这些特性与以0.2或50 mM的离子强度发生核裂解无关。考虑到体内的染色质组织和NHP迁移的可能作用，认为互补的可溶性和不溶性染色质组分之间的NHP含量差异很大。

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