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酵母REV1蛋白的脱氧基转移酶活性

J R Nelson¹, C W Lawrence, D C Hinkle

¹Department of Biophysics, University of Rochester, New York 14642-8408, USA.

|August 22, 1996

概括

在Saccharomyces cerevisiae中的REV1蛋白表现出脱氧代转移酶活性,插入细胞因子与DNA基底部位相反. 这种作用对于DNA聚合酶-zeta (Pol-zeta) 介导的DNA损伤后的转化合成至关重要.

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:

在Saccharomyces cerevisiae中的DNA损伤突变发生依赖于REV1,REV3和REV7基因.
Rev3和Rev7是DNA聚合酶-泽塔 (Pol-zeta) 的子单元,对于转化合成至关重要.
Rev1与大肠杆菌的UmuC具有同质性,通过不清楚的机制参与转化合成.

研究的目的:

为了研究Rev1蛋白的酶活性.
阐明Rev1在DNA损伤耐受性和突变发生中的作用.
了解Rev1和Pol-zeta在DNA修复途径中的相互作用.

主要方法:

在试验室中评估净化Rev1蛋白的脱氧基转移酶活性.
使用依赖模板的DNA原始延伸试验.
通过不同的DNA聚合酶 (Pol-zeta和Pol-alpha) 来表征原料延伸效率.

主要成果:

Rev1蛋白催化了从dCTP转移到DNA原料的3'末端的脱氧胺单酸盐 (dCMP) 残留的模板依赖转移.
有效的dCMP插入发生在模板基础站点对面.
显著的,虽然较小,插入发生在对面的模板关氨酸,腺氨酸和 uracil 基.
基底位相对的细胞因子插入创建了一个由Pol-zeta有效扩展的初始终端,但不是由酵母Pol-alpha.

结论:

Rev1具有一种新型的脱氧基转移酶活性,对DNA损伤突变发生至关重要.
这种活动促进了对面的基底部位的细胞因子的插入,从而使得Pol-zeta.通过后续扩展.
Rev1的功能是转化合成途径的组成部分,特别是绕过DNA损伤.

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