McLennan A G, Keir H M
Biochim Biophys Acta. 1975 Oct 15;407(3):253-62. doi: 10.1016/0005-2787(75)90092-1.
The subcellular location and growth stage dependence of the DNA polymerases of Euglena gracilis strain Z and of a bleached derivative of the strain have been studied by fractionation of the enzymes from extracts of whole cells and subcellular fractions on DEAE-cellulose. A new method for the rapid isolation of nuclei was employed. Of the major enzymes, pol A has a predominantly nuclear location and pol B a predominantly cytoplasmic location. Pol A is 4-fold and pol B 15-fold more active in exponentially-growing cells than in stationary-phase cells, pol B representing 90% of the combined activities in exponential-phase cells. The activity of the mitochondrial DNA polymerase increases about 3-fold as the cells enter stationary phase while that of the chloroplast DNA polymerase is greater in exponential-phase cells. The chloroplast enzyme persists in cells which have been reversibly bleached. The results are compared to those of similar experiments involving primitive and higher eucaryotes.
通过在二乙氨基乙基纤维素上对来自全细胞提取物和亚细胞组分的酶进行分级分离,研究了纤细裸藻Z菌株及其漂白衍生物的DNA聚合酶的亚细胞定位和生长阶段依赖性。采用了一种快速分离细胞核的新方法。在主要的酶中,聚合酶A主要定位于细胞核,聚合酶B主要定位于细胞质。在指数生长期细胞中,聚合酶A的活性是稳定期细胞的4倍,聚合酶B的活性是稳定期细胞的15倍,在指数生长期细胞中,聚合酶B占总活性的90%。当细胞进入稳定期时,线粒体DNA聚合酶的活性增加约3倍,而叶绿体DNA聚合酶在指数生长期细胞中的活性更高。叶绿体酶在可逆漂白的细胞中持续存在。将这些结果与涉及原始真核生物和高等真核生物的类似实验结果进行了比较。