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首页> 外文期刊>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：大鼠肝细胞核的非组蛋白蛋白及其核酸酶消化产生的染色质级分的分布

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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°C with or without nuclease and a further wash step. Nuclei at this stage had a remarkably constant NHP content (ca. 0.73 μg/μg 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 μg NHP/μg 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：在维持高阶染色质结构中的寻找蛋白质已经导致在核酸酶消化研究的背景下对大鼠肝细胞核的非组蛋白蛋白（NHP）进行了系统的检查。 40-45％的 3 h-therptophan标记的NHP最初存在的NHP可以通过在“生理学”缓冲液中进行大量洗涤，在37℃下孵育或不含核酸酶和进一步的洗涤步骤。在该阶段的核具有显着恒定的NHP含量（约0.73μg/μgDNA），与微球体核酸酶或HAEIII的消化程度无关。通过用核酸酶的有限消化产生的溶解的染色质含有0.3-0.5μg/μgDNA，该值落到CA. 0.16次裂解后。不溶性染色质级分在NHP中的2倍（消化物）和16倍（广泛的消化）中比相应的可溶性级分在NHP之间。凝胶电泳显示可溶性级分中约12个NHP带，最突出的M.Wt。 41.400，而不溶性材料具有至少50个组分。这些性能与核的裂解在0.2或50mM离子强度下发生。在体内染色质组织方面考虑了互补溶解和不溶性染色质级分之间的NHP含量的巨大视差，以及NHP迁移的可能作用。

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