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含有转录活性基因和无活性基因的鸡红细胞染色质结构域的溶解性和结构

Solubility and structure of domains of chicken erythrocyte chromatin containing transcriptionally competent and inactive genes.

作者信息

Komaiko W, Felsenfeld G

出版信息

Biochemistry. 1985 Feb 26;24(5):1186-93. doi: 10.1021/bi00326a020.

Abstract

Chromatin generated by micrococcal nuclease digestion of erythrocyte nuclei can be fractionated into two pools of differing solubility in solvents containing 0.15-0.25 M NaCl. A fixed percentage of the chromatin is soluble under these conditions, independent of the average size of the DNA in the unfractionated chromatin. Chromatin containing particular gene sequences is also distributed between soluble and insoluble fractions in a way that is independent of the average size of the starting material. However, the actual percentage of gene copies present in each fraction is not necessarily the same as for bulk chromatin. The transcriptionally active chicken erythrocyte adult beta-globin gene is more soluble than the bulk, while the ovalbumin gene in the same tissue is less soluble. These differences do not appear to be related to variations in content of RNA, core histones, or two classes of non-histone proteins. Instead, we find that the soluble chromatin pool is somewhat depleted in histones H1 and H5 and contains lower molecular weight DNA than precipitable chromatin. The soluble fraction can be made insoluble by addition of H1. If the precipitable chromatin fraction is redigested to reduce its size and then recombined with the soluble fraction and reprecipitated, the distribution of globin gene is randomized. The results suggest that the partitioning of chromatin into soluble and insoluble pools in 0.15-0.25 M NaCl arises from redistribution of a limiting amount of histones H1 and H5 to the chromatin fractions containing the longest DNA.

摘要

通过微球菌核酸酶消化红细胞核产生的染色质,在含有0.15 - 0.25M氯化钠的溶剂中可分为两个溶解度不同的部分。在这些条件下,一定比例的染色质是可溶的,这与未分级染色质中DNA的平均大小无关。含有特定基因序列的染色质也以一种与起始材料平均大小无关的方式分布在可溶和不可溶部分之间。然而,每个部分中基因拷贝的实际百分比不一定与整体染色质相同。转录活跃的鸡红细胞成人β - 珠蛋白基因比整体染色质更易溶,而同一组织中的卵清蛋白基因则较难溶。这些差异似乎与RNA、核心组蛋白或两类非组蛋白的含量变化无关。相反,我们发现可溶染色质部分中组蛋白H1和H5有所减少,并且与可沉淀染色质相比含有分子量更低的DNA。通过添加H1可使可溶部分变得不可溶。如果将可沉淀染色质部分再次消化以减小其大小,然后与可溶部分重新组合并再次沉淀,珠蛋白基因的分布就会随机化。结果表明,染色质在0.15 - 0.25M氯化钠中分为可溶和不可溶部分,是由于有限量的组蛋白H1和H5重新分布到含有最长DNA的染色质部分所致。

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