Remodeling and Repositioning of Nucleosomes in Nucleosomal Arrays.

Remodeling and Repositioning of Nucleosomes in Nucleosomal Arrays.
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核小体阵列中核小体的重塑和重新定位

DOI:
10.1007/978-1-4939-8556-2_18
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发表时间:
2018
影响因子:
--
通讯作者:
Mueller-Planitz
Mueller-Planitz
中科院分区:
--
文献类型:
--
作者:
Ludwigsen;Klinker;Pfennig;Mueller-Planitz

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atp依赖性核小体重塑因子塑造真核染色质的核小体景观。它们在称为核小体滑动的过程中沉积或排出核小体或沿着DNA重新定位核小体。传统上用单核小体作为模型底物来分析重塑。然而,在体内,核小体形成具有规则间距的扩展阵列。在这里,我们描述了如何在体外重建规则间隔的核小体阵列,以及如何将这些阵列用于解剖试管中的重构。我们概述了两个试验。第一种检测方法感知到核小体阵列内特定核小体的各种结构变化,而第二种检测方法则特异于检测阵列内核小体的重新定位。这两种分析方法都利用了重塑反应中DNA对限制性内切酶的可及性的变化。
ATP-dependent nucleosome remodeling factors sculpt the nucleosomal landscape of eukaryotic chromatin. They deposit or evict nucleosomes or reposition them along DNA in a process termed nucleosome sliding. Remodeling has traditionally been analyzed using mononucleosomes as a model substrate. In vivo, however, nucleosomes form extended arrays with regular spacing. Here, we describe how regularly spaced nucleosome arrays can be reconstituted in vitro and how these arrays can be used to dissect remodeling in the test tube. We outline two assays. The first assay senses various structural changes to a specific nucleosome within the nucleosomal array whereas the second assay is specific toward detecting repositioning of nucleosomes within the array. Both assays exploit changes to the accessibility of DNA to restriction enzymes during the remodeling reaction.
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发表时间: 2012-04-01
影响因子: 7
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