HsMCM8/9直酶的活性,基质偏好和结构
David R McKinzey1, Chuxuan Li2, Yang Gao2
1Department of Chemistry and Biochemistry, Baylor University, Waco, TX 76706, USA.
Nucleic acids research
|June 13, 2023
概括
人类MCM8/9 (微染色体维护8/9) 蛋白质作为依赖ATP的DNA螺旋体起作用. 它们的结构和机制揭示了一种对DNA复制叉稳定性和修复至关重要的顺序转位过程.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 遗传学 遗传学 是一个
背景情况:
- MCM8和MCM9是最近进化的MCM蛋白质,在较高的真核生物中发现.
- 在MCM8/9中发生的突变与不孕不育,卵巢衰竭和癌症有关.
- 已知MCM8/9在DNA复制和修复中的作用,但其生物化学活性和结构尚不清楚.
研究的目的:
- 为了阐明人类MCM8/9 (HsMCM8/9) 的生物化学活性和结构.
- 了解通过HsMCM8/9.9解开DNA的机制.
主要方法:
- 生物化学分析以确定DNA酶活性和基质特异性.
- 低温电子显微镜 (cryo-EM) 用于确定HSMCM8/9异构相的结构.
- 核酸结合研究.核酸结合研究.
主要成果:
- HsMCM8/9 作为一种依赖ATP的DNA螺旋酶,具有3'-5'极性.
- 高亲和度单链DNA结合发生在核酸三酸盐中,通过ATP水解减弱.
- 冷-EM结构显示了一种具有明显AAA+核酸结合点的异构体的三元体.
- 核酸结合诱导AAA+域的结构变化和显著的域移动.
结论:
- HsMCM8/9使用一个顺序子单元转位机制来解DNA.
- 结构和生物化学数据为MCM8/9在DNA复制和修复中的功能提供了机械洞察力.
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