Ebralidse K K, Grachev S A, Mirzabekov A D
Institute of Molecular Biology, Academy of Sciences, Moscow, USSR.
Nature. 1988 Jan 28;331(6154):365-7. doi: 10.1038/331365a0.
A nucleosomal core particle is composed of two each of histones H2A, H2B, H3 and H4 located inside the particle with approximately 47 base pairs (bp) of DNA wrapped around the octamer in about 1.8 turns of a left-handed superhelix. The path of the superhelix is not smooth; the DNA is sharply bent, or kinked, at positions symmetrically disposed at a distance of about one and four double-helical turns in both directions from the nucleosomal dyad axis (designated as sites +/- 1 and +/- 4 respectively). This non-uniform bending is considered archetypal to other DNA-protein complexes, but its mechanism is not clear (reviewed in ref. 4). DNA-histone chemical cross-linking within the core particle has revealed strong binding of each of the two histone H4 molecules to DNA at a distance of 1.5 helical turns either side of the nucleosomal dyad axis (sites +/- 1.5). In each of these sites, a single flexible domain of H4 was previously shown to contact three points, at about nucleotides 55 and 65 on one strand and nucleotide 88 on the complementary strand, numbering from the 5' terminus of each 147-base strand; these three locations are closely juxtaposed across the highly compressed minor and major grooves (Fig. 1). Here we report that the amino-acid residue of histone H4 cross-linked at the 1.5 site is histidine-18, embedded in a highly basic cluster Lys-Arg-His-Arg-Lys-Val-Leu-Arg which is probably involved in the sharp bending of the DNA double helix at the +/- 1 sites.
核小体核心颗粒由位于颗粒内部的组蛋白H2A、H2B、H3和H4各两个组成,约47个碱基对(bp)的DNA以左手超螺旋约1.8圈缠绕在八聚体周围。超螺旋的路径并不平滑;DNA在距核小体二分轴两侧约一个和四个双螺旋圈对称分布的位置处急剧弯曲或扭结(分别指定为位点+/-1和+/-4)。这种不均匀弯曲被认为是其他DNA-蛋白质复合物的典型特征,但其机制尚不清楚(参考文献4综述)。核心颗粒内的DNA-组蛋白化学交联揭示了两个组蛋白H4分子中的每一个在核小体二分轴两侧1.5个螺旋圈的距离处与DNA有强结合(位点+/-1.5)。在这些位点中的每一个,先前已表明H4的单个柔性结构域与三个点接触,从每条147个碱基链的5'末端开始编号,一条链上约为核苷酸55和65,互补链上为核苷酸88;这三个位置在高度压缩的小沟和大沟中紧密相邻(图1)。在此我们报告,在1.5位点交联的组蛋白H4的氨基酸残基是组氨酸-18,它嵌入一个高度碱性的簇Lys-Arg-His-Arg-Lys-Val-Leu-Arg中,该簇可能参与了DNA双螺旋在+/-1位点处的急剧弯曲。