酵母DNA聚合酶ε的 [4Fe4S] 集群具有逆氧活性,可以接受DNA介导的信号传递
Miguel N Pinto1, Josy Ter Beek2, Levi A Ekanger1,3
1Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, United States.
Journal of the American Chemical Society
|September 24, 2021
概括
DNA聚合酶 ε (Pol ε) 使用 [4Fe4S] 集群进行复制. 氧化会使酶失活,但是再减少,可能是通过DNA电荷传输,恢复活性.
科学领域:
- 生物化学
- 分子生物学
- 生物有机化学
背景情况:
- 许多DNA复制和修复酶使用4Fe4S集群.
- 酵母DNA聚合酶 ε (Pol ε) 的催化域,Pol2CORE,包含一个 [4Fe4S] 集群.
研究的目的:
- 在Pol2CORE中研究4Fe4S集群的氧化还原特性.
- 确定聚合氧化和还原对Pol ε活动的影响.
主要方法:
- 使用DNA电化学研究氧化还原特性.
- 采用了缺异核酶的Pol2CORE变体 (Pol2COREexo-) 和一个 [4Fe4S] 缺乏集群的突变.
- 通过使用DNA修复酶EndoIII检查了再减少的效果.
主要成果:
- 氧化Pol2COREexo-呈现出阴极氧化还原信号,而没有在4Fe4S无突变体中.
- 蛋白质氧化显著降低了聚合活性.
- 重制恢复了聚合酶活性,而EndoIII则通过DNA电荷传输促进了这种恢复.
结论:
- 在 Pol ε 中的 [4Fe4S] 集群具有氧化还原活性,并影响聚合酶功能.
- DNA电荷传输可以调节从修复酶到Pol ε的电子传输,调节其活性.
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