Russell W C, Precious B, Martin S R, Bayley P M
EMBO J. 1983;2(10):1647-53. doi: 10.1002/j.1460-2075.1983.tb01639.x.
The right-handed (B) conformation of poly[d(G-C)] in 7.5 mM sodium cacodylate and 25% ethylene glycol can be readily converted to the left-handed (Z) conformation by the addition of 250 microM MnCl2 and this transition can be reversed by chelation of the Mn ions with EDTA or by addition of NaCl. This ability to obtain such reversible transitions in solvent and solute conditions which allow DNA-protein interactions and their assessment by c.d. permitted an analysis of the effect of purified histones, polyamino acids, protamine and polyamines on these transitions. Individual core histones H3, H4, H2a and H2b or protamine stabilised the Mn-induced Z form and prevented the transition to B DNA normally observed after chelation with EDTA or on dialysis to physiological salt concentrations. A similar suppression of Z leads to B transition was also achieved with poly-L-arginine (but not with poly-L-lysine). In contrast, histones H1 and H5 promoted the Z leads to B transition. Polyamines (spermine and spermidine) converted the B form to another right-handed (A) form which transformed to the Z form after the addition of EDTA and this Z form was restored to the B conformation on the addition of NaCl. These results suggest that sequence-dependent variations in the conformation of natural DNA may be modulated by interaction with histones and other basic cellular components and may provide a conformational basis for nucleosome formation and possibly for the control of gene expression.
在7.5 mM 二甲胂酸钠和25%乙二醇中,聚[d(G-C)]的右手(B)构象可通过添加250 microM 氯化锰轻松转化为左手(Z)构象,并且通过用EDTA螯合锰离子或添加氯化钠可使这种转变逆转。在允许DNA与蛋白质相互作用并通过圆二色性进行评估的溶剂和溶质条件下获得这种可逆转变的能力,使得能够分析纯化的组蛋白、聚氨基酸、鱼精蛋白和多胺对这些转变的影响。单个核心组蛋白H3、H4、H2a和H2b或鱼精蛋白可稳定锰诱导的Z型构象,并防止在用EDTA螯合后或透析至生理盐浓度时通常观察到的向B型DNA的转变。聚-L-精氨酸(但聚-L-赖氨酸不行)也实现了对Z向B转变的类似抑制。相比之下,组蛋白H1和H5促进了Z向B的转变。多胺(精胺和亚精胺)将B型转化为另一种右手(A)型,在添加EDTA后这种A型转变为Z型,并且在添加氯化钠后这种Z型恢复为B构象。这些结果表明,天然DNA构象中依赖序列的变化可能通过与组蛋白和其他基本细胞成分的相互作用来调节,并且可能为核小体形成以及可能为基因表达的控制提供构象基础。