Smirnov I V, Dimitrov S I, Makarov V L
Institute of Molecular Biology Academy of Science of the USSR, Moscow.
J Biomol Struct Dyn. 1988 Apr;5(5):1149-61. doi: 10.1080/07391102.1988.10506455.
We have used flow linear dichroism (LD) and light scattering at 90 degrees to study the condensation of both DNA and calf thymus chromatin by polyamines, such as spermine, spermidine and its analogs designated by formula NH3+(CH2)iNH2+(CH2)jNH3+, where i = 2,3 and j = 2,3, putrescine, cadaverine and MgCl2. It has been found that the different polyamines affect DNA and chromatin in a similar way. The level of compaction of the chromatin fibers induced by spermine, spermidine and the triamines NH3+(CH2)3NH2+(CH2)3NH3+ and NH3+(CH2)3NH2+(CH2)2NH3+ and MgCl2 is found to be identical. The triamine NH3+(CH2)3NH2+(CH2)2NH3+ and the diamines studied condense neither chromatin nor DNA. This drastic difference in the action of the triamines indicates that not only the charge, but also the structure of the polycations might play essential roles in their interactions with DNA and chromatin. It is shown that a mixture of mono- and multivalent cations affect DNA and chromatin condensation competitively, but not synergistically, as claimed in a recent report by Sen and Crothers (Biochemistry 25, 1495-1503, 1986). We have also estimated the extent of negative charge neutralization produced by some of the polyamines on their binding to chromatin fibers. The stoichiometry of polyamine binding at which condensation of chromatin is completed is found to be two polyamine molecules per DNA turn. The extent of neutralization of the DNA phosphates by the histones in these compact fibers is estimated to be about 55%. The model of polyamine interaction with chromatin is discussed.
我们利用流动线性二色性(LD)和90度光散射来研究多胺(如精胺、亚精胺及其通式为NH3+(CH2)iNH2+(CH2)jNH3+的类似物,其中i = 2,3且j = 2,3)、腐胺、尸胺和MgCl2对DNA和小牛胸腺染色质的凝聚作用。已发现不同的多胺对DNA和染色质的影响方式相似。精胺、亚精胺、三胺NH3+(CH2)3NH2+(CH2)3NH3+、NH3+(CH2)3NH2+(CH2)2NH3+以及MgCl2诱导的染色质纤维压缩程度相同。三胺NH3+(CH2)3NH2+(CH2)2NH3+以及所研究的二胺既不凝聚染色质也不凝聚DNA。这些三胺作用的显著差异表明,多阳离子的电荷以及结构在它们与DNA和染色质的相互作用中可能都起着至关重要的作用。结果表明,单价和多价阳离子的混合物对DNA和染色质凝聚的影响是竞争性的,而非如Sen和Crothers最近的报告(《生物化学》25, 1495 - 1503, 1986)中所声称的协同作用。我们还估计了一些多胺与染色质纤维结合时产生的负电荷中和程度。发现染色质凝聚完成时多胺结合的化学计量比为每DNA螺旋两分子多胺。在这些紧密纤维中,组蛋白对DNA磷酸根的中和程度估计约为55%。文中讨论了多胺与染色质相互作用的模型。