Wallace P G, Hewish D R, Venning M M, Burgoyne L A
Biochem J. 1971 Nov;125(1):47-54. doi: 10.1042/bj1250047.
regenerating rat liver, normal rat liver, rat thymus, normal mouse liver and Ehrlich ascites-tumour cells. In all cases two main polymerase forms are observed, one of mol.wt. 200000, preferring denatured DNA to native calf thymus DNA primer, designated type I, and the other, designated type II, of mol.wt. 100000, showing a variable and slight preference for native calf thymus DNA primer. Some catalytic properties of these polymerases are described. Nuclei have been isolated from some of these tissues by using two different buffer systems. The ionic composition of the isolation medium is found to affect greatly the amounts and types of polymerase that bind to the nuclei, and also affects the kinetic properties of the polymerases. The way the polymerases and nuclei change properties as the ionic composition of the buffers is changed suggests that ionic effects may be a significant factor in the control of DNA synthesis in vivo. These ionic effects also explain much of the previous confusion over the localization of specific DNA polymerases.
对以下真核生物组织的DNA聚合酶进行了研究:再生大鼠肝脏、正常大鼠肝脏、大鼠胸腺、正常小鼠肝脏和艾氏腹水肿瘤细胞。在所有情况下,均观察到两种主要的聚合酶形式,一种分子量为200000,与天然小牛胸腺DNA引物相比,更倾向于变性DNA,称为I型;另一种称为II型,分子量为100000,对天然小牛胸腺DNA引物表现出可变且轻微的偏好。描述了这些聚合酶的一些催化特性。通过使用两种不同的缓冲系统,从其中一些组织中分离出了细胞核。发现分离介质的离子组成极大地影响与细胞核结合的聚合酶的数量和类型,并且还影响聚合酶的动力学特性。随着缓冲液离子组成的变化,聚合酶和细胞核性质的改变方式表明,离子效应可能是体内DNA合成控制中的一个重要因素。这些离子效应也解释了之前在特定DNA聚合酶定位方面的许多困惑。