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分离染色质亚基的DNA分析

Analysis of DNA of isolated chromatin subunits.

作者信息

Lacy E, Axel R

出版信息

Proc Natl Acad Sci U S A. 1975 Oct;72(10):3978-82. doi: 10.1073/pnas.72.10.3978.

Abstract

Partial digestion of rat liver nuclei with staphylococcal nuclease results in the liberation of nucleo-protein complexes consisting of one or more upsilon bodies. By velocity centrifugation we have isolated the monomeric subunit in relatively pure form. We find that this subunit contains 185 base pairs of DNA and 240,000 daltons of protein, resulting in a protein to DNA ratio identical to that of unperturbed chromatin. The isolated monomeric particle is further susceptible to internal nuclease attack resulting in the solubilization of 46% of the monomeric DNA. Analysis of the resistant DNA reveals a complex but highly reproducible pattern of DNA fragments ranging from 160 to 60 base pairs in length. Analysis of the reassociation kinetics of the isolated subunit DNA reveals that most, if not all genomic sequences, are involved in this basic subunit structure. No special frequency class of DNA is absent from upsilon bodies. Furthermore, virtually all liver sequences transcribed into mRNA are present in upsilon body DNA. These results indicate that upsilon body formation may be random with respect to DNA sequence and suggest that the mere presence of upsilon bodies over a specific region of DNA is not sufficient to restrict its transcription.

摘要

用葡萄球菌核酸酶对大鼠肝细胞核进行部分消化,会释放出由一个或多个υ体组成的核蛋白复合物。通过速度离心,我们已分离出相对纯的单体亚基。我们发现该亚基含有185个碱基对的DNA和240,000道尔顿的蛋白质,其蛋白质与DNA的比例与未受干扰的染色质相同。分离出的单体颗粒更容易受到内部核酸酶的攻击,导致46%的单体DNA溶解。对抗性DNA的分析揭示了长度从160到60个碱基对的复杂但高度可重复的DNA片段模式。对分离出的亚基DNA的复性动力学分析表明,大多数(如果不是全部)基因组序列都参与了这种基本亚基结构。υ体中不存在特殊频率类别的DNA。此外,几乎所有转录成mRNA的肝脏序列都存在于υ体DNA中。这些结果表明,υ体的形成可能与DNA序列无关,并且表明仅在DNA的特定区域存在υ体不足以限制其转录。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/090f/433121/7ec6324f40d3/pnas00053-0197-a.jpg

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