人类DNA聚合酶 μ 可用于多个Mn2+-介导功能的非正规机制
Yao-Kai Chang1,2, Ya-Ping Huang1, Xiao-Xia Liu1
1Institute of Biological Chemistry, Academia Sinica , 128 Academia Road Sec. 2 , Nankang, Taipei 115 , Taiwan.
Journal of the American Chemical Society
|May 9, 2019
概括
DNA聚合酶mu (Pol μ) 使用非正规的预结合途径用于金属结合的核酸,显示对而不是的偏好. 这种机制对于其在DNA修复和复制中的多样性功能至关重要.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- DNA聚合酶是DNA复制和修复的关键酶.
- 在一些DNA聚合酶中观察到一种非正规的途径,涉及金属结合核酸 (MdNTP) 的预结合.
- DNA聚合酶mu (Pol μ) 的主要途径和金属偏好,X家族成员具有独特的Mn2+偏好,仍然不清楚.
研究的目的:
- 调查非正规的预结合途径是否是Pol μ.的主要机制.
- 阐明Polμ不寻常的Mn2+偏好的结构基础.
- 了解这种途径在各种Polμ函数中的作用.
主要方法:
- 使用X射线晶体学进行结构分析.
- 确定了19个结构,包括预结合的二进制,前催化三进制和产品复合体.
- 生物化学测试以评估核酸结合和金属离子偏好.
主要成果:
- 聚米先将MdNTP结合在一种催化活性构造中,支持一种非正规的通路.
- 在多个功能的预结合阶段,Pol μ表现出Mn2+对Mg2+的偏好.
- 结构数据揭示了非正典机制和Mn2+偏好的基础.
结论:
- 非正规的预结合途径在功能上与Polμ的不同作用有关.
- 这项研究解决了Polμ的Mn2+偏好的长期难题.
- 这些发现扩大了已知的DNA聚合酶的机械景观.
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