High-affinity binding sites for histone H1 in plasmid DNA.
High-affinity binding sites for histone H1 in plasmid DNA.
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质粒 DNA 中组蛋白 H1 的高亲和力结合位点。
DOI：
10.1073/pnas.92.15.7060
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发表时间：
1995
影响因子：
11.1
通讯作者：
Zlatanova,J
中科院分区：
文献类型：
--
作者：
Yaneva,J;Schroth,GP;vanHolde,KE;Zlatanova,J
用凝胶电泳法研究了鸡红细胞组蛋白H1与质粒pBR322和pUC19限制性内切酶片段的相互作用。某些限制片段对组蛋白表现出异常高的亲和力，在蛋白质与DNA的比率下形成高分子量复合体，而其他片段没有显示出结合的证据。高度优选的片段本质上是弯曲的，根据它们在聚丙烯酰胺凝胶中的电泳迁移率、计算机模拟和扫描力显微镜成像来判断。然而，用相同片段的弯曲部分或高度弯曲的动泡体DNA片段进行的对照实验表明，仅有曲率的存在不足以进行优先结合。通过使用以高度优先的序列为中心的各种限制片段，发现高亲和力结合另外需要在曲率区域两侧存在特定序列。因此，曲率和特定位点的存在似乎都需要产生高亲和力。
The interaction of histone H1 isolated from chicken erythrocytes with restriction fragments from plasmids pBR322 and pUC19 was studied by gel electrophoresis. Certain restriction fragments exhibited unusually high affinity for the histone, forming high molecular mass complexes at protein to DNA ratios at which the other fragments did not show evidence for binding. The highly preferred fragments are intrinsically curved, as judged by their electrophoretic mobility in polyacrylamide gels, by computer modeling, and by imaging with scanning force microscopy. However, control experiments with either curved portions of the same fragments or highly curved kinetoplast DNA fragments showed that the presence of curvature alone was not sufficient for preferential binding. By using various restriction fragments centered around the highly preferred sequence, it was found that the high-affinity binding required in addition the presence of specific sequences on both sides of the region of curvature. Thus, both curvature and the presence of specific sites seem to be required to generate high affinity.