Camerini-Otero R D, Sollner-Webb B, Felsenfeld G
Cell. 1976 Jul;8(3):333-47. doi: 10.1016/0092-8674(76)90145-8.
We have examined the role played by various histones in the organization of the DNA of the nucleosome, using staphylococcal nuclease as a probe of DNA conformation. When this enzyme attacks chromatin, a series of fragments evenly spaced at 10 base pair intervals is generated, reflecting the histone-DNA interactions within the nucleosome structure. To determine what contribution the various histones make to DNA organization, we have studied the staphylococcal nuclease digestion patterns of complexes of DNA with purified histones. Virtually all possible combinations of homogeneous histones were reconstituted onto DNA. Exhaustive digestion of a complex containing the four histones H2A, H2B,H3, and H4 yields a DNA fragment pattern very similar to that of whole chromatin. The only other combinations of histones capable of inducing chromatin-like DNA organization are H2A/H2B/H4 and those mixtures containing both H3 and H4. From an examination of the kinetics of digestion of H3/H4 reconstitutes, we conclude that although the other histones have a role in DNA organization within the nucleosome, the arginine-rich histone pair, H3/H4, can organize DNA segments the length of the nucleosome core in the absence of all other histones.
我们利用葡萄球菌核酸酶作为DNA构象的探针,研究了各种组蛋白在核小体DNA组织中所起的作用。当这种酶攻击染色质时,会产生一系列以10个碱基对间隔均匀分布的片段,这反映了核小体结构内组蛋白与DNA的相互作用。为了确定各种组蛋白对DNA组织的贡献,我们研究了DNA与纯化组蛋白复合物的葡萄球菌核酸酶消化模式。实际上,所有可能的纯合组蛋白组合都被重组到了DNA上。对含有四种组蛋白H2A、H2B、H3和H4的复合物进行彻底消化,产生的DNA片段模式与整个染色质的模式非常相似。能够诱导类似染色质的DNA组织的唯一其他组蛋白组合是H2A/H2B/H4以及同时含有H3和H4的混合物。通过对H3/H4重组体消化动力学的研究,我们得出结论,虽然其他组蛋白在核小体的DNA组织中起作用,但富含精氨酸的组蛋白对H3/H4在没有所有其他组蛋白的情况下也能组织核小体核心长度的DNA片段。