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在S. cerevisiae中的第三个必需的DNA聚合酶

A Morrison¹, H Araki, A B Clark

¹Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences Research Triangle Park, North Carolina 27709.

|September 21, 1990

概括

基因聚合酶I和III对于酵母的生存至关重要. 这项研究表明,DNA聚合酶II也是必不可少的,挑战了当前的真核生物DNA复制模型.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物化学生物化学

背景情况:

众所周知,DNA聚合酶I和III对于Saccharomyces cerevisiae (酵母菌) 的生存能力至关重要.
在酵母复制中,其他核DNA聚合酶 (如DNA聚合酶II) 的作用和必要性仍然不太清楚.

研究的目的:

克隆和分析POL2基因,该基因编码了酵母DNA聚合酶II的催化子单元.
确定DNA聚合酶II对酵母细胞活力和DNA复制的必要性.

主要方法:

在S. cerevisiae中克隆和分析POL2的基因.
转录分析以确定POL2mRNA的大小.
蛋白质序列分析以预测分子重量和识别功能域.
基因破坏和删除实验,以评估对细胞活力和DNA聚合酶活性的影响.

主要成果:

克隆了POL2基因,发现它表达了一个编码为255,649Mr蛋白的7.5kb的转录.
对POL2的部分破坏导致了具有截断DNA聚合酶II活性的可活细胞.
完全删除POL2读取框架导致了不可行性和特征性的DNA复制停止形态.
预测的蛋白质的N终端区域与其他真核细胞DNA聚合酶具有有限的序列相似性.

结论:

三种核DNA聚合酶 (I,II和III) 对于S. cerevisiae的生存能力至关重要.
DNA聚合酶II在DNA复制中发挥着关键作用,可能作为复制酶起作用.
这三种DNA聚合酶作为复制酶的本质性挑战了现有的真核DNA复制模型.

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