Selective displacement of histone H1 from whole HeLa nuclei: effect on chromatin structure in situ as probed by micrococcal nuclease.
Selective displacement of histone H1 from whole HeLa nuclei: effect on chromatin structure in situ as probed by micrococcal nuclease.
复制标题
组蛋白 H1 从整个 HeLa 核中的选择性置换：微球菌核酸酶探测对原位染色质结构的影响。
DOI：
10.1021/bi00578a005
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发表时间：
1979
期刊：
影响因子：
2.9
通讯作者：
R. Cole
中科院分区：
文献类型：
--
作者：
G. Lawson;R. Cole
George M. Lawson*和R. David Cole*摘要：为了在最大限度地减少染色质变性的条件下研究组蛋白HI在染色质结构中的作用，开发了一种从整个细胞核中置换组蛋白的技术。分离的HeLa细胞核在pH 3.0下选择性地释放组蛋白HI，核心组蛋白H2A， H2B， H3和H4没有检测到移位。通过分别用微球菌核酸酶和胰腺DNA酶I原位酶切产生的双链DNA片段的特征性电泳带带模式来判断，洗涤后的核小体结构保持完整。这些模式与维持生理ph的细胞核产生的模式相同。含有组蛋白HI的对照细胞核或完全或部分失去组蛋白HI的细胞核中，核小体核之间的核溶裂动力学都明显是一级的，每种情况下都可以用单一速率常数来描述。组蛋白HI定量置换后，总切割率增加了2.7±0.2倍。定量反比关系
George M. Lawson* and R. David Cole* abstract: In order to study the role of histone HI in chromatin structure under conditions minimizing chromatin denaturation, a technique was developed to displace this histone from whole nuclei. Isolated HeLa nuclei washed at pH 3.0 selectively released histone HI without detectable displacement of the core histones H2A, H2B, H3, and H4. Nucleosomal structure remained intact in such washed nuclei as judged by the characteristic electrophoretic banding patterns of dou-ble-stranded DNA fragments generatedby in situ digestion with micrococcal nuclease and pancreatic DNase I, respec-tively. The patterns were identical with those produced from nuclei maintained at physiological pH. The kinetics of nu-cleolytic cleavage of the linkage DNA between nucleosome cores in control nuclei containing histone HI or in nuclei completely or partially depleted of histone HI were all ap-parently first order and in each case could be described with a single rate constant. The overall rate of cleavage was in-creased by a factor of 2.7±0.2 following quantitative displacement of histone HI. A quantitative inverse relationship