Hecht N B
Biochim Biophys Acta. 1975 Apr 2;383(4):388-98. doi: 10.1016/0005-2787(75)90308-1.
After aqueous subcellular fractionation and partial purification by phosphocellulose chromatography, murine cells are found to contain a low molecular weight DNA-dependent DNA polymerase (beta) in the nuclear fraction and two distinguishable DNA-dependent DNA polymerases (C-I and C-II) in the cytosol. Both C-I and C-II are found in testis, liver, and regenerating liver; the amount of C-I being several fold increased in the regenerating liver and in immature testis. C-I and C-II are distinguishable by the criteria of salt sensitivity, inhibition by single-stranded DNA, elution from phosphocellulose, inhibition by 0.3 mM N-ethylmaleimide, template preference, and sedimentation coefficient. C-II is dissociated by 0.25 M KC1 to an active form of DNA polymerase of sedimentation coefficient 3.5 S while C-I is not dissociated, maintaining its sedimentation coefficient of 7.2 S. Many similar chemical and physical properties of C-II and the low molecular weight nuclear DNA polymerase (beta) suggest that C-II may represent an aggregate state of beta monomers, The size, reaction properties and the increase in enzyme activity under conditions of rapid cellular proliferation suggest C-I is analogous to the alpha DNA polymerase.
经水相亚细胞分级分离及磷酸纤维素层析部分纯化后,发现鼠细胞的核部分含有一种低分子量的依赖DNA的DNA聚合酶(β),而胞质溶胶中含有两种可区分的依赖DNA的DNA聚合酶(C-I和C-II)。C-I和C-II在睾丸、肝脏及再生肝中均有发现;再生肝和未成熟睾丸中C-I的含量增加了几倍。C-I和C-II可通过盐敏感性、单链DNA抑制作用、从磷酸纤维素上的洗脱、0.3 mM N-乙基马来酰亚胺的抑制作用、模板偏好性及沉降系数等标准加以区分。C-II可被0.25 M KCl解离为沉降系数为3.5 S的活性DNA聚合酶形式,而C-I则不会解离,其沉降系数保持为7.2 S。C-II与低分子量核DNA聚合酶(β)的许多相似化学和物理性质表明,C-II可能代表β单体的聚集状态。C-I的大小、反应特性以及在细胞快速增殖条件下酶活性的增加表明它类似于α DNA聚合酶。