Fapy.dG指示Klenow exo(-) 错误地将脱氧氨基素纳入其中
Carissa J Wiederholt1, Marc M Greenberg
1Department of Chemistry, Colorado State University, Ft. Collins, Colorado 80523, USA.
N-(2-deoxy-alpha,beta-d-erythropentofuranosyl) -N-(2,6-diamino-4-hydroxy-5-formamidopyrimidine) (Fapy.dG) 显著增加了克莱诺的DNA复制错误. 这种损伤导致频繁的错误整合和低效的扩展,导致错误率高出800万倍.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 遗传学 是一个遗传学.
背景情况:
- 在复制过程中,DNA损伤会导致突变.
- 了解特定病变如何影响聚合酶活性对于DNA修复和突变性研究至关重要.
研究的目的:
- 调查Fapy.dG对Klenow外聚合酶活性的影响.
- 量化Fapy.dG在DNA合成过程中的易出错性.
主要方法:
- 合成了含有特定部位Fapy.dG病变的寡核酸基质.
- 使用这些基质评估了Klenow外聚合酶活性.
- 测量了核酸结合和原料扩展效率.
主要成果:
- 在两个关键步骤中,Fapy.dG 抑制了原料聚合:在病变的对面加入,并在病变的过去延伸.
- 与dG相比,Klenow exo-表现出与Fapy.dG相对应的脱氧腺酸错误结合的频率高出20倍.
- 用Fapy.dG:dA对扩展一个原料,效率只比用Fapy.dG:dC对稍低一点.
结论:
- 与dG相比,Fapy.dG显著损害了Klenow的异常保真性,使总体错误概率增加了大约8000万倍.
- Fapy.dG是一种强大的突变性病变,它促进了错误的结合和通过DNA聚合酶的绕过.
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