人类DNA聚合酶 δ的冷-EM结构阐明了它在没有PCNA的情况下最小的DNA合成活性
Yeonoh Shin1, Mark Hedglin2, Katsuhiko S Murakami1
1Department of Biochemistry and Molecular Biology, The Center for RNA Molecular Biology, The Center for Structural Biology, The Pennsylvania State University, University Park, Pennsylvania, USA.
人类DNA聚合酶三角酶 (Pol δ) 需要增殖细胞核抗原 (PCNA) 来实现高DNA合成活性. 一个新发现的形Pol δ的酸螺旋抑制了DNA结合,这解释了它在没有PCNA的情况下的低活性.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 生物化学 生物化学
背景情况:
- DNA聚合酶三角酶 (Pol δ) 对于真核生物的DNA复制和修复至关重要.
- 人类Pol δ在没有增殖细胞核抗原 (PCNA) 的情况下表现出最小的活动,但原因尚不清楚.
研究的目的:
- 阐明PCNA依赖人类Pol δ活动的结构基础.
- 了解管理Pol δ功能的监管机制.
主要方法:
- 使用冷电子显微镜 (cryo-EM) 确定了阿波形人类Pol δ. 的结构.
- 结构分析的重点是Pol δ子单元和潜在的DNA结合部位之间的相互作用.
主要成果:
- 人类Pol δ (p125,p66,p50,p12) 的阿波形体的冷-EM结构在3.65 Å时得到解决.
- 发现p125亚单元中的一个N端酸性α螺旋体占据了DNA结合腔.
- 这种螺旋可能在没有PCNA的情况下抑制DNA结合,解释了低的apo-form活性.
结论:
- 人类Pol δ中的酸性α-螺旋体作为一种自身抑制的元素,解释了PCNA依赖.
- 在酵母Pol δ中缺少这种螺旋体,可以解释PCNA独立活性中特定物种的差异.
- 这些发现为波兰的监管及其对PCNA的依赖提供了关键的见解.
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