Simpson R T
J Biol Chem. 1979 Oct 25;254(20):10123-7.
The well separated transitions observed on heating synthetic core particles containing poly(dA-dT).poly(dA-dT) have allowed physicochemical studies to define the mechanism of the first, reversible transition. The shape of the particle, assessed by sedimentation velocity measurements, becomes extended at temperatures about 10 degrees C below the optical melting temperature; this change is unaffected by cross-linking the histones into a stable octamer. A portion of the DNA undergoes a transition to a structure more like that of protein-free poly(dA-dT).poly(dA-dT), as judged by circular dichroism measurements, also at temperatures below those necessary for optical melting. Digestion of 5'-32P-labeled particles with S1 nuclease at a temperature partly through the first optical transition demonstrates that the initial phase in melting core particles is separation of about 20 base pairs of DNA at each end of the particle segment from the histone core. Implications of this mechanism for transcription of histone-bound DNA segments in chromatin are discussed.
在加热含有聚(dA-dT)·聚(dA-dT)的合成核心颗粒时观察到的明显分离的转变,使得物理化学研究能够确定第一次可逆转变的机制。通过沉降速度测量评估的颗粒形状,在比光学熔解温度低约10摄氏度的温度下会变得伸展;这种变化不受将组蛋白交联成稳定八聚体的影响。通过圆二色性测量判断,一部分DNA也在低于光学熔解所需温度的情况下转变为更类似于无蛋白聚(dA-dT)·聚(dA-dT)的结构。在部分经历第一次光学转变的温度下用S1核酸酶消化5'-32P标记的颗粒表明,熔解核心颗粒的初始阶段是颗粒片段两端约20个碱基对的DNA与组蛋白核心分离。讨论了这种机制对染色质中组蛋白结合DNA片段转录的影响。